SNP (Single Nucleotide Polymorphism) molecular marker related to economic traits of Pinan cattle and application of SNP molecular marker

Abstract

The invention discloses an SNP (Single Nucleotide Polymorphism) molecular marker related to economic traits of Pinan cattle and application of the SNP molecular marker. Through whole genome re-sequencing (WGRS), whole genome association analysis (GWAS), transcriptome function annotation and linkage imbalance analysis, a key single nucleotide polymorphism (SNP) site rs135687714 (exon region missense mutation) of an APLF gene is identified in Pican-south cattle, and the site and rs137691885 (an intron region, a QTL marker site) disclosed in cattle QTL have medium linkage (R2 = 0.5). Genetic typing and character correlation analysis show that individuals with different genotypes of rs135687714 have significant differences (Plt; 0.01) in 11 growth characters such as body width, hip width, hoof and foot development and the like. The molecular marker constructed based on the APLF gene SNP can be used for pionan cattle germplasm identification and molecular assisted selection (MAS), efficient identification and breeding can be carried out in the early stage of calves, an efficient, simple, convenient and stable technical means is provided for molecular breeding of beef cattle, and the molecular marker has important application value and popularization prospects.

Description

Technical Field

[0001] This invention relates to the fields of molecular biology, genomics, and transcriptomics, specifically to a SNP molecular marker associated with economic traits in Pinan cattle and its application. Background Technology

[0002] Using whole-genome resequencing technology, over 8100 SNP variants related to growth traits in the Pinan cattle genome were comprehensively captured. Combined with whole-genome sequence information, these SNPs were rapidly located to specific genes or gene regions. Genome-wide association analysis (GWAS) is a powerful tool for annotating phenotypic effects on the genome. By analyzing growth-trait-related genes and chromosomal regions in cattle, it can efficiently improve the selection and breeding of beef cattle genomes and genes. Bioinformatics methods were used to perform quantitative analysis, differential expression analysis, and corresponding gene function annotation and enrichment analysis on the extensive genomic data collected, identifying the roles of specific genes in biological regulation in the Pinan cattle population and revealing gene regulatory functions. In the field of animal breeding, the selection of DNA markers closely related to growth traits improves breeding accuracy.

[0003] However, current research on the functional genes and molecular markers of economic traits in Pinan cattle is limited, especially regarding the function of DNA repair-related genes in beef cattle development. Therefore, there is an urgent need to screen for stable functional SNP markers to improve the accuracy and efficiency of molecular breeding. Summary of the Invention

[0004] The present invention aims to solve the above-mentioned technical problems by providing a method and application for selecting molecular markers related to the growth traits of Pinan cattle.

[0005] To solve the above-mentioned technical problems, the technical solution provided by the present invention is as follows: A SNP molecular marker associated with economic traits in Pinan cattle, the molecular marker being located in the exon region of the APLF gene on bovine chromosome 11, specifically at the rs135687714 site, where the base is mutated from T to C, causing a missense mutation of the amino acid from Met to Thr.

[0006] The molecular marker is linked to the rs108964133 locus on chromosome 11 in the cattleQTL database (R²=0.5), and the two can be used as a combined molecular marker for Pinan cattle breeding.

[0007] The PCR primer pair for the SNP molecular marker associated with the economic traits of Pinan cattle has the following primer sequences: SEQ ID NO.2 (forward primer): 5'-CAGGACTGCTTGAAGATGGA-3'; SEQ ID NO.3 (reverse primer): 5'-TGGTGTTCTCAGCCTCTTTC-3'.

[0008] The aforementioned KASP primer pair for SNP molecular markers related to the economic traits of Pinan cattle includes: SEQ ID NO.4 (Allele T-specific primer, FAM tail): 5'-GAAGGTGACCAAGTTCATGCT–TCTCAGGACTGCTTGAAGATGGT-3'; SEQ ID NO.5 (allele C-specific primer, HEX tail): 5'-GAAGGTCGGAGTCAACGGATT–TCTCAGGACTGCTTGAAGATGGC-3'; SEQ ID NO.6 (Universal reverse primer): 5'-AGCAGGAAGGACACTGACAC-3'.

[0009] A molecular detection method using primer pairs, wherein the molecular detection method can perform genotyping of the rs135687714 locus in individual Pinan cattle, for use in MAS-assisted breeding.

[0010] The application of an SNP molecular marker or detection method in early molecular-assisted selection of Pinan cattle to identify individuals associated with 11 growth traits, including hip width, height, and body structure.

[0011] The technical approach of this invention includes, but is not limited to, the following key steps: 2.1 Sample Collection and Multi-omics Data Integration: Distant bulls were selected from the core breeding population of Pinan cattle, and fresh venous blood samples were collected to extract whole-genome DNA. A sequencing library was constructed, and 30X whole-genome sequencing was completed. This data was then combined with whole-genome data from 22 other beef cattle breeds in public databases (see appendix). Figure 1 ), and conduct population genetic structure and nucleotide diversity analysis.

[0012] 2.2 SNP variation and positive selection analysis: A triple selective clearance analysis using the population differentiation index (Fst), nucleotide diversity ratio (θπ), and neutral validation index (Tajima's D) was conducted to identify highly confident positively selected regions in the Pinan cattle population. Significantly differentiating missense mutation sites were screened by comparing allele frequencies between Pinan cattle and 22 other beef cattle populations.

[0013] 2.3 Functional Notes: Gene enrichment analysis is used to interpret the functions or specific features that are significantly enriched in a genome or collection of genes. The positive selection gene enrichment analysis of Pinan cattle and 22 other cattle breeds showed that significantly enriched gene entries and pathways included 1p36 copy number variation syndrome (WP5345), membrane organization (GO:0061024), 4p16 3 copy number variation (WP5365), embryonic development ending in birth or egg hatching (GO:0009792), monoatomic cation transmembrane transport (GO:0098655), growth (GO:0040007), reproductive structure development (GO:0048608), and heart development (GO:0007507). These key biological processes involve complex gene regulatory networks and participate in multi-level gene regulation by encoding specific proteins. (Appendix) Figure 2 ).

[0014] 2.4 Chain Imbalance Analysis: The APLF gene (aprataxin and PNKP-like factor) is primarily involved in DNA repair, participating in a process called non-homologous end joining (NHEJ). When DNA breaks occur during replication, the protein encoded by the APLF gene helps to reconnect the broken ends, ensuring the normal transmission of genetic information within the cell. Studies have shown that mutations in the APLF gene may cause anemia in humans, while its function in Pinan cattle remains unclear. The APLF gene is located on bovine chromosome 11, region 66,865,005-66,958,571, with ENSEMBLE ID ENSBTAG00000018401.

[0015] In Pinan cattle, a missense mutation at the rs135687714 site in the APLF gene coding region causes one amino acid to be replaced by another in the protein, altering the type of amino acid encoded. The mutation frequency at this site is 85%. (See appendix) Figure 3Genome-wide association analysis (GWAS) results showed that the mutation at the rs108964133 site of the APLF gene, located in the 1-2 intron region, significantly affects the growth, development, body structure, and other traits of cattle, and can be used as an important molecular marker in the field of breeding.

[0016] Association analysis between positively selected APLF gene expression loci in the Pinan cattle population and loci published in Cattle QTL revealed a moderate linkage (R²=0.5) between the rs135687714 locus in the APLF gene coding region and the rs108964133 locus in the Cattle QTL database. This linkage suggests that APLF can serve as a co-molecular marker, jointly influencing the growth and development of cattle, thus confirming the important molecular marker function of the APLF gene in the Pinan cattle population.

[0017] 2.5 Protein structure prediction and functional analysis: Homology modeling and the SWISS-MODEL protein structure prediction platform were used to assess the impact of key amino acid variations on the three-dimensional structure of encoded proteins. By combining hydrophobic and hydrophilic physicochemical parameters and the spatial relationship between variant sites and conserved domains, the influence of missense mutation sites in the APLF gene in the Pinan cattle population on the function of expressed proteins was identified, thereby enhancing the accuracy of this study.

[0018] In Pinan cattle, a missense mutation T>C in the exon 7 region of the APLF gene causes the encoded amino acid 276 to change from Met to Thr, resulting in a change in the protein structure of the APLF gene and causing α-helix deletion. (See appendix) Figure 4 ).

[0019] The hydrophilicity and hydrophobicity of proteins are crucial determinants of their spatial folding and functional realization. The coding region of the APLF gene at locus rs135687714 and adjacent regions exhibit significantly enhanced hydrophilicity (see appendix). Figure 5 This affects the surface properties of proteins.

[0020] 2.6 Analysis of typical functional SNP sites in Pinan cattle: Compared with existing technologies, this invention has the following beneficial effects: By analyzing positively selected genes related to growth and developmental traits in the Pinan cattle genome, this invention discovered a linkage relationship between the APLF gene exon rs135687714 and intron rs108964133, which jointly affect the development of economic traits in Pinan cattle. Furthermore, mutations at APLF gene exon sites alter the structure of the protein encoded by this gene, enhancing its hydrophilicity and affecting the characteristics of the APLF gene-expressed protein, thereby enhancing the DNA repair function of the Pinan cattle population.

[0021] 2.7 Application Potential: Given the crucial role of the APLF gene in the development of economic traits in Pinan cattle, the rs135687714 and rs108964133 loci were combined and applied to molecular-assisted selection (MAS) to predict growth performance in calves early on. Furthermore, it can be developed into a chip molecular marker library and extended to large-scale population screening and breeding.

[0022] 2.8 Beneficial effects: This invention clarifies for the first time the functional association between the APLF gene rs135687714 locus and growth traits in Pinan cattle, and verifies its genetic effect using Cattle QTL. rs135687714 is a functional mutation site, and rs108964133 is a previously reported QTL marker site. The linkage between the two (R² = 0.5) can jointly improve typing accuracy and application stability. This molecular marker can be rapidly detected in early beef cattle breeding, significantly improving the breeding efficiency of the core Pinan cattle population.

[0023] This invention also relates to a method for selecting molecular markers associated with growth traits in Pinan cattle, comprising the following steps: Sample collection and multi-omics data integration: Fresh venous blood samples were collected from the core breeding population of Pinan cattle, whole-genome DNA was extracted, a sequencing library was constructed, and high-throughput resequencing was performed; genomic data of other beef cattle breeds and RNA-seq datasets related to beef cattle growth traits were downloaded from public databases. SNP variation and positive selection analysis: Bioinformatics software was used to compare, detect variations, perform quality control, and annotate the genome data of Pinan cattle and other local beef cattle breeds to obtain a high-quality SNP locus dataset. Multiple screening methods, including population differentiation index, nucleotide diversity ratio, and neutral validation index, were used to identify multiple SNP mutation sites of genes related to growth traits in the Pinan cattle population. Transcriptome differential expression and functional enrichment analysis: By combining transcriptome data and functional annotation analysis, a differential gene expression matrix was constructed, and intersection analysis was performed with the genes containing candidate SNP sites to identify candidate genes that were co-expressed and upregulated. GO functional annotation and enrichment analysis were then performed on the set of candidate genes carrying functional mutations. Protein structure-function prediction: Homology modeling methods and related platforms were used to evaluate the impact of key amino acid variations on the three-dimensional structure of proteins. Combined with hydrophobic and hydrophilic physicochemical parameters and the spatial relationship between variant sites and conserved domains, the impact of mutation sites on protein function was evaluated, and molecular markers related to the growth traits of Pinan cattle were screened.

[0024] Preferably, in step (1), the number of fresh venous blood samples collected from 10 Pinan cattle is 10, and the DNBSEQ-T7 platform is used for paired-end 150bp high-throughput resequencing; the genome data of other beef cattle breeds downloaded are 342, and the RNA-seq dataset is 10.

[0025] Preferably, in step (2), the bioinformatics software includes BWA, GATK, VCFtools, and VEPtool; the population differentiation index is Fst, the nucleotide diversity ratio is θπ, and the neutrality verification index is Tajima's D.

[0026] Preferably, in step (3), 10 published beef cattle muscle tissue transcriptome datasets are selected from the NCBIGEO database to construct a differential gene expression matrix.

[0027] Preferably, in step (4), the relevant platform is the SWISS-MODEL platform.

[0028] Preferably, the molecular markers selected that are associated with the growth traits of Pinan cattle are SNP sites on the LRP2 gene, including c.2363G>C site, c.2617C>T site, c.5471A>G site, c.6259A>G site, c.8377A>C site, c.9706T>C site, c.10079T>A site, and c.11519G>A site.

[0029] Preferably, the LRP2 gene is located on bovine chromosome 2 and participates in biological processes such as cell signaling, cell adhesion, and metabolic regulation.

[0030] Another aspect of this invention relates to the application of screened molecular markers in the assessment of genetic diversity, germplasm resource evaluation, and genetic breeding of Pinan cattle populations.

[0031] Preferably, the molecular markers are used to construct SNP chips and develop KASP and TaqMan specific molecular markers to achieve early prediction and precise selection of the growth performance of individual Pinan cattle.

[0032] Given the crucial role of the LRP2 gene in muscle development in Pinan cattle, the findings of this invention are of great significance for constructing SNP chips for molecular-assisted selection (MAS), developing specific molecular markers such as KASP and TaqMan, and screening core breeding bulls in Pinan cattle. Specifically, it has the following beneficial effects: This invention is the first to jointly analyze whole-genome resequencing data of Pinan cattle with public transcriptome data, achieving cross-scale integration from population-level variant identification to molecular functional-level expression verification and structural analysis, thereby improving the accuracy and reliability of screening results.

[0033] We systematically screened target genes with multi-site mutations (such as LRP2) from candidate genes. Specifically, we used methods such as hydrophobicity change prediction, protein spatial conformation modeling, and conserved domain localization to verify the potential biological effects of SNPs, providing a solid theoretical foundation for subsequent molecular marker development.

[0034] Intersection analysis of upregulated genes co-expressed across multiple transcriptomes significantly improves the accuracy and biological explanatory power of functional SNP site screening, overcoming the shortcomings of traditional markers that are based solely on population frequencies.

[0035] The study found that key SNP variations (such as c.2363G>C and c.8377A>C in the LRP2 gene) can lead to changes in protein hydrophobicity and perturbations of key structural domains. This revealed their impact on nutrient transport and protein reabsorption at the molecular level, which is directly related to the rapid growth trait of Pinan cattle, providing new targets for the genetic breeding of Pinan cattle.

[0036] Candidate functional SNP sites have clear functional annotations, physical locations, and sequence information, which can be used to develop high-throughput genotyping technologies such as KASP and TaqMan, construct a molecular toolbox for the germplasm evaluation of local cattle breeds, realize early prediction and precise selection of the growth performance of individual Pinan cattle, accelerate the breeding process of superior breeding cattle, reduce breeding costs, and improve breeding efficiency. Attached Figure Description

[0037] Figure 1 This is a map showing the location of the LRP2 gene c.2363G>C and c.8377A>C sites in exon 16 and exon 44 of chromosome 2. Figure 2 This is a comparison diagram of the differences between the protein structure of the LRP2 gene before mutation at the c.2363G>C site (Wild-type, WT) and after mutation (Mutant, Mut). Figure 3 This is a comparison diagram of the differences between the protein structure of the LRP2 gene before and after the mutation (Mutant, Mut) at the c.8377A>C site in this invention. Figure 4 The diagram shows that the hydrophilic and hydrophobic properties of the protein in this invention are important determinants of its spatial folding and functional realization.

[0038] Figure 5 This is a genome information diagram of Pinan cattle and other beef cattle of the present invention.

[0039] Figure 6 This is a pathway entry diagram showing the significant enrichment of positively selected genes in Pinan cattle according to the present invention.

[0040] Figure 7 This invention relates to the exon rs135687714 of the APLF gene in Pinan cattle and its linked intron rs108964133, as well as the allele frequency map of the APLF gene in Pinan cattle and other beef cattle breeds.

[0041] Figure 8 This is a comparative diagram showing the differences between the protein structure (A) before the mutation at the rs135687714 site of the APLF gene (Wild-type, WT) and the protein structure (B) after the mutation (Mutant, Mut). (The red area represents the protein structure change caused by the amino acid mutation at the mutation site).

[0042] Figure 9 This is a comparison of the hydrophilicity and hydrophobicity of the APLF gene protein before and after the rs135687714 mutation. (The target protein was analyzed using the ProtScale tool, and the solubility tendency of each amino acid residue was quantified based on the Kyte-Doolittle hydrophobicity scale. The green line represents the hydrophilicity and hydrophobicity of the protein before the mutation, and the red line represents the hydrophilicity and hydrophobicity of the protein after the mutation. The APLF gene rs135687714 mutation enhances the hydrophilicity of the protein.) Detailed Implementation

[0043] The present invention will now be described in further detail with reference to the full text.

[0044] Combined with appendix Figures 1-9 , Example 1: Sample Collection and Sequencing Distant individuals from the core population of Pinan cattle were collected and subjected to 30× whole-genome resequencing. The population genetic structure and nucleotide diversity were compared with publicly available genomic data from other beef cattle breeds.

[0045] Example 2: GWAS and Chaining Analysis: In the Cattle QTL, the APLF gene rs108964133 locus was significantly associated with body depth (P < 0.05), hip width (P < 0.05), and foot and leg development (P < 0.05). Linkage analysis showed that the linkage disequilibrium coefficient between the APLF gene rs108964133 and rs135687714 loci was R² = 0.5, indicating a linkage reaction between the two SNP loci.

[0046] Example 3: Effects of APLF gene SNP site mutations on protein structure Using bioinformatics analysis tools such as Swiss-Model, ProtScale, PolyPhen-2, and SIFT, protein structure modeling and hydrophilicity / hydrophobicity prediction were performed on SNP mutations in the coding region of the APLF gene. The results showed that the mutation at the rs135687714 site of the APLF gene caused missense mutations and changes in protein structure, resulting in increased protein hydrophilicity.

[0047] Example 4: PCR typing Amplification was performed using SEQ ID NO.2 / 3, with a product length of approximately 230 bp. Genotype frequencies were determined using Sanger sequencing.

[0048] Example 5: KASP Classification Genotyping using SEQ ID NO.4-6 clearly distinguished three genotypes with an accuracy of 99%.

[0049] Example 6: Application Calves were genotyped at 3 months of age, and their body development potential was predicted based on rs135687714. Compared with traditional family selection, breeding efficiency was increased by approximately 20%, and the selection cycle for core breeding bulls was shortened by 6-8 months.

[0050] This invention also provides a method for screening functional mutation sites closely related to the rapid growth phenotype of Pinan cattle using whole-genome resequencing and transcriptomics data, employing methods such as population genetic analysis, selection signal detection, differential expression analysis of growth traits, gene function enrichment, and protein structure-function localization analysis. Furthermore, it involves analyzing the protein structure, hydrophobicity, and protein function of multiple SNP sites in the same gene (such as the c.2363G>C site and c.8377A>C site in the LRP2 gene) to verify their potential functional effects. Subsequently, it establishes a molecular marker-assisted breeding system based on these key mutations, enabling early prediction and precise selection of individual Pinan cattle growth performance, accelerating the breeding process of superior breeding cattle, and promoting the development of the local beef cattle industry. This invention relates to the fields of molecular biology, genomics, and transcriptomics. Specifically, it uses a multi-omics approach combining whole-genome resequencing and transcriptome expression analysis to screen molecular markers (SNP sites) of functional genes significantly related to beef cattle growth traits, and applies them to the fields of Pinan cattle population genetic diversity assessment, germplasm resource evaluation, and genetic breeding.

[0051] The technical approach of this invention includes the following key steps: (1) Sample collection and multi-omics data integration: Fresh venous blood samples (n=10) were collected from the core breeding population of Pinan cattle, and whole-genome DNA was extracted to construct sequencing libraries. Paired-end 150bp high-throughput resequencing was performed using the DNBSEQ-T7 platform. Genomic data of 342 other local beef cattle breeds and 10 publicly available RNA-seq datasets related to beef cattle growth traits were downloaded from public databases for transcriptome differential analysis.

[0052] Specifically, the genomic information of 342 beef cattle was compared: the domestic herd includes 67 cattle of 6 breeds (Kazakh, Tibetan, Lingnan, Wenshan, Yunnan, and Xiangxi); the foreign herd includes 275 cattle of 16 breeds (Angus, Red Angus, Hereford, Jersey, Holstein, Charolais, Griffith, Simmental, Korean, Shorthorn, Anchorage, Brahman, Ndama, Bagaria, Semien, and Bale). See below for details.

[0053] Domestic beef: The genome accession numbers for Kazakh in Xinjiang are: SRR5507260, SRR5507259, SRR5507258, SRR5507257, SRR5507256, SRR5507255, SRR5507254, SRR5507253, and SRR5507252.

[0054] The Tibetan genome accession numbers are: SRR5507251, SRR5507250, SRR6024571, SRR5507249, SRR6024572, SRR5507248, SRR5507247, SRR5507246, SRR5507245, SRR5507244, and SRR5507243.

[0055] The genome accession numbers for Lingnan in Shaanxi are: SRR5507222, SRR5507221, SRR5507220, SRR5507219, SRR5507218, SRR5507217, SRR5507216, and SRR5507215.

[0056] The genome accession numbers for Wenshan in Yunnan are: SRR6024561, SRR6024562, SRR6024569, SRR6024570, SRR6024577, SRR6024578, SRR6024575, and SRR6024576.

[0057] The genome accession numbers for Dianzhong in Yunnan are: SRR6024567, SRR6024568, SRR6024565, SRR6024566, SRR6024563, SRR6024564, SRR12739619, SRR12739618, SRR12739617, and SRR12739616.

[0058] The genome accession numbers for Xiangxi in Hunan are: SRR16949978, SRR16949979, SRR16949980, SRR16949981, SRR16949982, SRR16949983, SRR16949984, SRR16949985, SRR16949986, SRR16949987, and SRR16949. 988、SRR16949989、SRR16949990、SRR16949991、SRR16949992、SRR16949993、SRR1694999 4. SRR16949995, SRR16949996, SRR16949997, SRR16949998, SRR16949999, SRR16950000.

[0059] Foreign beef: Angus cattle from Aberdeenshire, Scotland, UK, with genome accession numbers: SRR1525582, SRR1525583, SRR1525686, SRR1525687, SRR1525688, SRR1355237, SRR1365144, SRR1425124, SRR1365129, SRR1346376, SRR866424, SRR1343174, SRR1346379, SRR1346391, SRR1348573, SRR1348580, SRR1348581, SRR135 5244, SRR1355255, SRR1355257, SRR1365132, SRR1425147, SRR1425148, SRR1425149, SRR1425150, SRR1365128, SRR1365131, SRR1365137, SRR 1348590, SRR1365105, SRR1365107, SRR1365109, SRR1365110, SRR1365111, SRR1365139, SRR1462188, SRR1462189, SRR1346381, SRR1346380.

[0060] The Red Angus cattle from Aberdeenshire, Scotland, UK, have the following genome accession numbers: SRR1525580, SRR1525581, SRR1525600, SRR1525613, SRR1525614, SRR1343172, SRR1355239, SRR1365103, SRR1365113, SRR1425153, SRR1348588, SRR1348591, SRR1425145, SRR1425146, SRR1425152, and SRR1343170.

[0061] Hereford cattle in the UK, with genome accession numbers: SRR1343160, SRR1346382, SRR1346383, SRR1346385, SRR1346387, SRR1365124, SRR1365126, SRR1365128, SRR1365131, SRR1365137, SRR1348590, SRR1365105, SRR1365107, SRR1365109, SRR1365110, SRR1365111, SRR1365139, SRR1462188, SRR1462189, SRR1346381, SRR1346380.

[0062] Jersey cattle in Channel Island of Jersey, UK, have the following genome accession numbers: SRR3497161, SRR3497162, SRR3497451, SRR3497462, SRR3497464, SRR3497465, SRR3497466, SRR3497467, SRR3497611, SRR3497613, SRR2016774, and SRR2016776.

[0063] Holstein cattle from DeKooy, North Holland, Holland, with genome accession numbers: SRR1346386, SRR1346390, SRR1346392, SRR1348583, SRR1348592, SRR1365147, SRR1425129, SRR1425133, SRR1425134, SRR1425144, SRR1365101, SRR1365104, SRR1365114, SRR1365115, SRR1365116, SRR1365117, SRR1365118, SRR1365119, SRR1365120, SRR1365121, SRR 1425125, SRR1425126, SRR1425127, SRR1425128, SRR1425130, SRR1425131, SRR14 25132, SRR1425135, SRR1425136, SRR1425137, SRR1425138, SRR1425139, SRR1425 155. SRR1425156, SRR1425157, SRR1425158, SRR1425159, SRR1425160, SRR142516 1. SRR1425162, SRR1425163, SRR1425164, SRR1425165, SRR1425166, SRR1346389.

[0064] Charolais cattle from France, with genome accession numbers: SRR1343167, SRR1343168, SRR1343169, SRR1348571, SRR1355258, SRR1365122, SRR1343173, SRR1348574, SRR1348577, SRR1348578, SRR1348584, SRR1355253, SRR1355254, SRR1365136 The Gelbvieh cattle from München, Bavaria, Germany have the following genome accession numbers: SRR134316, SRR1343162, SRR1343164, SRR1343165, SRR1343166, SRR1365112, SRR1425154, SRR1355236, SRR1355240, SRR1355260, SRR1346388, SRR1355241, SRR1355242, SRR1355243, SRR1355246, SRR1355248, SRR1355249, SRR1355250, SRR1355251, and SRR1365138.

[0065] Geneve, a Simmental breed from Switzerland, has the following genome accession numbers: SRR1525617, SRR1525618, SRR1525619, SRR1525620, SRR1525621, SRR1525700, SRR1525701, SRR1525702, SRR1525703, SRR1525705, SRR1525622, SRR1525623, SRR1525625, SRR1525626, SRR1525627, SRR1525713, SRR1525714, SRR1525715, SRR1525716, SRR1525717, SRR1525718, and SRR1525719. The Hanwoo cattle from Chuncheon, Kangwon-do, Korea have the following genome accession numbers: SRR934415, SRR934417, SRR934418, SRR934419, SRR934432, SRR934433, SRR934434, SRR934435, SRR934437, SRR934395, SRR934397, SRR934398, SRR934400, SRR934401, SRR934402, SRR934403, and SRR934404.

[0066] The genome accession numbers for the Shorthorn Zebu cattle from Uganda (Closestcity) are: ERR320241, ERR320244, ERR320246, ERR320249, ERR320252, ERR320261, ERR320242, ERR320253, ERR320257, and ERR320254.

[0067] Ankole cattle from Uganda, Africa, with genome accession numbers: ERR320247, ERR320264, ERR320245, ERR320251, ERR320255, SRR3656914, SRR3656915, SRR3656919, SRR3656920, SRR3656921, SRR3656922, SRR3656924, SRR3225527, SRR3656912, SRR3656917, ERR320258, ERR358484, ERR320262, ERR320243, ERR320248 The Brahman cattle Boran of Kenya, Africa, have the following genome accession numbers: SRR3508267, SRR3524756, SRR3524807, SRR3524810, SRR3546725, SRR3546727, SRR3546728, SRR3546729, SRR3546777, and SRR3546787.

[0068] The NDama cattle of Guinea, Africa, have the following genome accession numbers: SRR3693229, SRR3693373, SRR3693376, SRR3693378, SRR3693379, SRR3693419, SRR3693420, SRR3694478, and SRR3694578.

[0069] The genome accession numbers for Bagaria cattle in Ethiopia and Africa are: SRR13626465, SRR13626466, SRR13626493, SRR13626494, SRR13626496, SRR13626467, SRR13626468, SRR13626469, SRR13626495, and SRR13626497.

[0070] Semien cattle from Ethiopia and Africa have the following genome accession numbers: SRR13626471, SRR13626473, SRR13626478, SRR13626480, SRR13626481, SRR13626472, SRR13626474, SRR13626475, SRR13626477, and SRR13626479.

[0071] The genome accession numbers for Bale cattle from Ethiopia and Africa are: SRR13626483, SRR13626484, SRR13626485, SRR13626488, SRR13626489, SRR13626491, SRR13626482, SRR13626486, SRR13626490, and SRR13626492. (2) SNP variation and positive selection analysis: By combining the geographical distribution differences between the Pinan cattle population and other beef cattle populations, bioinformatics software (BWA, GATK, VCFtools, VEPtool, etc.) was used to compare, detect variations, perform quality control, and functional annotation of the genomic data of 352 cattle, obtaining a high-quality single nucleotide polymorphism (SNP) dataset. Through multiple screening using the population differentiation index (Fst), nucleotide diversity ratio (θπ), and neutral validation index (Tajima's D), highly confident positive selection gene regulatory regions in the Pinan cattle population were identified, as well as multi-SNP mutation sites of genes related to growth traits.

[0072] (3) Transcriptome differential expression and functional enrichment analysis: To further verify the functional association between candidate SNPs related to growth traits obtained through genome resequencing and the actual phenotypic development of beef cattle, this invention combines transcriptome data and functional annotation analysis to conduct multi-omics joint analysis. Ten published beef cattle muscle tissue transcriptome datasets were selected from the NCBIGEO database, and a differential gene expression matrix was constructed. Intersection analysis was performed with the genes containing the candidate SNPs to identify candidate genes co-regulated in multiple datasets. Furthermore, GO (GeneOntology) functional annotation and enrichment analysis were performed on the set of candidate genes carrying functional mutations. The LRP2 gene, also known as LDL receptor-associated protein 2, is a member of the low-density lipoprotein receptor family, located on bovine chromosome 2, and participates in various biological processes such as cell signaling, cell adhesion, and metabolic regulation. The LRP2 gene is enriched in several growth-related gene entries, such as: heart development (GO:0007507), kidney development (GO:0001822), aortic development (GO:0035904), and forebrain development (GO:0030900).

[0073] (4) Protein structure-function prediction: Homology modeling and the SWISS-MODEL platform were used to evaluate the impact of key amino acid variations on protein three-dimensional structure. Combined with hydrophobic and hydrophilic physicochemical parameters and the spatial relationship between variant sites and conserved domains, the impact of mutation sites on protein function was evaluated, thereby improving the accuracy of SNP sites with altered biological functions.

[0074] (5) Analysis of typical functional SNP sites in Pinan cattle: The LRP2 gene, also known as LDL receptor-associated protein 2, is a member of the low-density lipoprotein receptor family, located on chromosome 2, and participates in various biological processes such as cell signaling, cell adhesion, and metabolic regulation. LRP2 is widely distributed in polarized epithelial cells, with the highest expression in the kidneys. It is mainly distributed in the brush border of the proximal tubules and the cambium of glomerular epithelial cells, forming a transporter together with the intrinsic factor-vitamin B12 receptor (Cubilin). LRP2 plays an important physiological role in the reabsorption of proteins, trace elements, and nutrients in the body. In animal models, LRP2 knockout mice exhibit severe bone disease characterized by low bone mineral content, poor bone hardness, and increased osteoid surfaces.

[0075] In Pinan cattle, eight missense mutations were detected at SNP sites in the LRP2 gene: c.2363G>C, c.2617C>T, c.5471A>G, c.6259A>G, c.8377A>C, c.9706T>C, c.10079T>A, and c.11519G>A. Among these, mutations at sites c.2363 and c.8377 in two exons resulted in complete mutations in Pinan cattle, altering amino acids 788 and 2793 of the LRP2 gene, causing protein structural torsion and alteration (see appendix). Figure 2 and 3 ).

[0076] like Figure 2 As shown, the differences in protein structure between the LRP2 gene before (Wild-type, WT) and after (Mutant, Mut) mutations at the c.2363G>C site are compared. The red area represents the protein conformational changes caused by the amino acid mutation at the mutation site.

[0077] like Figure 3 As shown, the differences in protein structure between the LRP2 gene before mutation at the c.8377A>C site (Wild-type, WT) and after mutation (Mutant, Mut) are compared. The red area represents the protein conformational change caused by the amino acid mutation at the mutation site.

[0078] The hydrophobic and hydrophobic properties of proteins are crucial determinants of their spatial folding and functional realization. The hydrophobicity of the coding region and adjacent regions of the c.2363G>C site in the LRP2 gene is significantly increased (see appendix). Figure 4 A), while the amino acid encoded by the c.8377A>C site of the LRP2 gene and its adjacent residues show significantly enhanced hydrophilicity (see appendix). Figure 4 B).

[0079] like Figure 4 As shown, the hydrophilicity and hydrophobicity of proteins are crucial determinants of their spatial folding and functional realization. The target protein was analyzed using the ProtScale tool, and the solubility tendency of each amino acid residue was quantified based on the Kyte-Doolittle hydrophobicity scale. The results in the figure show the difference in hydrophilicity and hydrophobicity between proteins before and after mutations at the c.2363G>C and c.8377A>C sites in the LRP2 gene (Mutant, Mut). The green line represents the hydrophilicity and hydrophobicity of the protein before mutation, and the red line represents the hydrophilicity and hydrophobicity of the protein after mutation. The mutation at the c.2363G>C site in the LRP2 gene increases the hydrophobicity of the protein, while the mutation at the c.8377A>C site in the LRP2 gene increases the hydrophilicity of the protein.

[0080] Conserved domains are functional units that maintain high sequence similarity and structural stability during protein evolution. They typically correspond to specific catalytic active sites, ligand binding sites, or protein-protein interaction interfaces, performing specific biological functions and playing a crucial role in maintaining protein structure and function. Using CDD to predict the domains of the target protein, it was found that LRP2c.2363G>C is located in the LDL receptor repetitive sequence class B superfamily domain (LDL_recept_bsuperfamily). This repetitive sequence exists in multiple tandem repeats and is predicted to form a β-propeller structure. LRP2c.8377A>C is located in the LDL receptor domain; this cysteine-rich repetitive sequence in the receptor plays a central role in mammalian cholesterol metabolism.

[0081] Sample collection and genome resequencing: Ten core breeding individuals of Pinan cattle with complete growth trait records were selected, and peripheral blood samples were collected. High-quality genomic DNA was extracted using the TIANamp blood DNA extraction kit. After the DNA quality was tested and found to be satisfactory by Nanodrop and Qubit, an Illumina library was constructed and subjected to paired-end 150bp high-throughput sequencing on the MGIDNBSEQ-T7 platform, with an average sequencing depth of approximately 30×.

[0082] like Figure 1 As shown, the c.2363G>C and c.8377A>C sites of the LRP2 gene are located in exons 16 and 44 of chromosome 2, respectively. The figure also shows the allele frequency distribution of the LRP2 gene in the genomes of 10 Pinan cattle and 342 other beef cattle breeds (6 domestic and 16 foreign breeds) across different populations. The Pinan cattle breed maintains its unique allele mutations.

[0083] Whole-genome variation detection and annotation: After adapter removal and low-quality sequence filtering using FastP software, the raw sequencing data was cleaned and aligned to the Bostaurus reference genome (ARS-UCD1.2) using BWA. SNP and InDel variant detection was performed using GATK. VCFtools was used for further variant filtering, and VEP was used for SNP functional annotation. Approximately 81 million SNPs were obtained, of which 41.79 million were newly discovered variants not included in the dbSNP database.

[0084] Population structure and selection signal analysis: Combining sequencing data from Pinan cattle with resequencing data from 104 representative local cattle (such as Nanyang cattle, Luxi cattle, and Leizhou cattle) and introduced cattle (Angus and Simmental cattle) in publicly available databases, we used PCA, NJ tree, and ADMIXTURE to analyze population structure and genetic background. Using a combined analysis of F_ST, πratio, and Tajima's D methods, we identified multiple candidate regions regulated by positive selection, covering functional gene regions related to bone growth, muscle development, and oxidative stress.

[0085] Functional mining of candidate genes and SNPs: Multiple candidate genes associated with growth traits, such as GCLC, PARVA, NDN, FMN, LRP, and LGR, were screened within the selected signal region. Specifically, eight SNP missense mutations were found in exons 16 and 44 of the LRP2 gene: c.2363G>C, c.2617C>T, c.5471A>G, c.6259A>G, c.8377A>C, c.9706T>C, c.10079T>A, and c.11519G>A.

[0086] The effect of LRP2 gene SNP mutations on the structure and function of encoded proteins: Bioinformatics analysis tools such as Swiss-Model, ProtScale, PolyPhen-2, and SIFT were used to perform structural modeling, conserved domain prediction, and functional prediction of the wild-type (WT) and mutant (Mut) proteins encoded by the LRP2 gene. The results showed that there are 8 SNP mutations in the LRP2 gene. Mutations at the c.2363G>C and c.8377A>C sites caused changes in protein structure and hydrophobicity, thus affecting the surface properties of the encoded protein. Conserved domain prediction revealed that LRP2 c.2363G>C is located in the LDL_recept_bsuperfamily, a repeat sequence that forms a β-propeller structure, causing changes in protein structure. LRP2 c.8377A>C is located in the LDL_recept_bsuperfamily, a cysteine-rich repeat sequence that plays a central role in mammalian cholesterol metabolism.

[0087] Breeding applications and SNP molecular marker development: Based on multi-site mutations in the LRP2 gene, KASP primers were designed to validate genotyping in the core Pinan cattle population. Results showed that the C allele frequency was significantly higher in the Pinan cattle population than in other breeds, and individuals with this genotype exhibited superior performance in daily weight gain and carcass weight. Based on this mutation, KASP molecular markers were developed, and a MAS-assisted screening system was constructed for early breeding value assessment in calves and bull selection, significantly improving the breeding efficiency of Pinan cattle.

[0088] The amino acid sequence of the protein is shown below: CRSNEFLCQPGLCITASWRCDGTRDCPNGADEIDCPTSCHSNQFLCPNEQLCIPESWVC DGEDDCNDGADERRHCPGITCSSRQFTCENGECIPGEFRCDHSTDDCLDGTDEKNCRYPVC EQLTCADGACYNTSQRCNGQVDCRDASDERNCTHRCRTEFQCGSGQCIPRTYVCDHEID CEDGSDEHSCPAYRTCKGNEFTCPNGVCIAQSWVCDGESDCVDNADEDGCESKINRTFEC YPNEWACPKSGKCIPISKVCDGTLDCPGGEDESNITAGQQCDVNLCPSLGCEYQCHRSPG GGMCYCPSGFIVNQNRTNNCVDFDDCQIWGVCDQLCEDRIGHHRCNCVEGYVLERQKHCR ANSSFGEAFVIFSNGRNLLKGSIRGNNFQILAESQNRGLAVGVDFHYRLRRVFWTDIVQK KVFSVDISGSRIREVLGVSIEDPENLAVDWVNNKLYIVETRVNCIDVVDLDGSHRITLIS EYLGHPRGIAVDPTVGYLFFSDWQNVFGVPRIERAYMDGSNRKDLVTTKLGWPGGITLDL VSKRVYWVDSRFDYIETVTYDGLKRMTVIHGGADIPHPFSISLFEGQLFFTDWTKMAVLK ANKFKETNPRLYYRSSLKPFGVTVYHGLRQPYARNPCAHENGGCQHICVLSHRTFNGGLG YRCKCRLGYIPDWDDYHCVAAERFLLFSSNLAVRGIPLTLSHQTEVILPVTGSSSIFLGI DFDAREKAIFFSDTKKNIIYRQKLDGTGREIITANRVPAVQSLSFDWISRNLYWTDSSYR SVSVIRLGDKSRRTIIQNLNNPQSIVVHPTAGYIFFSVWYRPAKILRAWADGSNILPIVN TTLGWPSGLSIDWSSSRLYWVDAFFDKIEHSNLDGSDRKALTNVHQLTHPFGLAVSQDYI YVTDWRRGIIRFGKYNPGQSIILRSGVGSVMRAKVYDSRVQTGSNACSRPTNPNGDCSHF CFPVPNSQRVCGCPYGMSLASDHLTCVENASREPPVEQCGTLSFSCHNGRCVPLQYRCDG FDDCLDNSDEAQCTTSNATCSPLAFECKREGHCIPSMWRCDGEDDCLDGSDEQNCPTRAP TSCRADQFTCDNNFCIPRSWVCDTDNDCKDGSDEKSCNYTQTCSPTQFHCPDHRCIALTF VCDGTKDCADGSDEIGCVINCTASQFTCVSNGQCISKTYRCDGVFDCDDHSDETDCPTRP PGMCHQDEFQCQEDGICIPKTWECDGHEDCLQGSDEHNGCPPKTCHPSHFVCQNGNCIYR NWLCDGDNDCGDMSDEKDCPTQPFQCPSWQWQCPGHSICVNLSAVCDGVSDCPGGTDESP LCNQNSCSDSNGGCTHQCIQGPYGAQCQCPLGYLLGNDSKTCEDIDECRTPGFCSQYCYN MRGSFRCWCDSEYTLDADRRTCKATGNEKSETLLLVASQSQLVAGNMTQNGYFVYPVIQH GSHIVAVDFDSVSGRIFWSDGTQGKIWSAFQNGTDKKLILDSGVSMTGSIAVDWIGRNLY WTDISLRTIDVAKLDGSHKTVLISENITNLGGLAVDPRATDRVMFWSDWGSHPRIERASM DGSQRTIIVQEKIYWPNGLALDYPNRLLYFMDGYLDYLDFCHYDGSNRRQVIASDLILRH PYSISLFEDTVYWSDRATHEVMKANKWHGGNQSVVMTLHQPLGIVVVHPAKQPVSSNPCS YTRCSHLCLLSSKSLYSCACPSGWTLARDSVTCVRDDQAFLIVVRNSIIFGISLNPDVKT FDGMVPISGIRNGYDVAVDYSEQFIYWLENPGEIHRVKTDGTNRTVFAPLSSLGASASLA LDWLSRNLYFTDHVTRSIKVMTLQGDVSYRKTLIANDGTSLGVGLPVGITIDPINGKLYW SDRGTNSGLPPKIASANMDGKSPRTLFTGSLENVAFITLDIEEQKLYWAVSSTGVIERGN VDGTNRMILVNHLSYPWGLAVHGRFLYYSDEEYEVIERVDKATGANKVVLRNNLPDLRGL KIYQRRSESSNGCSNNMNACQQICLPVPGRLFSCACATGFKLNPDHQTCSPYNSFIVVST LRTIRGFSLQLSDHSEAMVPVAGPGRNALHVDVDVSSGFIYWCDFNISVASNNAIRRIKP DGSNFTNIVTDGIGVNGVRGIAVDWVAGNLYFTNAFRSETLIEVLRINTTHRHILLKTIV DMPRDIVVDPKNRYLFWSDYGQNPKIERSFLDCTNRTVLVSDITATPRGLALDHSSNYIY WVDDAVDLIARISIEGGETEVIRFGSHYPAPYAITVFGNSIIWVDRNLKKILQASKEPNR ADRPTVIRDNIDWLRDVTIFDQSVQPRSPAEVNNNPCLENNGGCAQFCFALPKSQTPKCD CAFGTLQADGKSCAISSENFLIFALDDSLRSLRFDPKDYSQPFPAISVERMAVALDYDSI DNRIYFTQLLPSGKGQISYINLNSRSSPPTVVVSGIGSPEGIAFDWINKRIYYSDYTNQM IKSIATDGSRHTLIAQVPKPRGIVLDPCQGYMYWTDWGTNAKIEMATMGGYSRRSLVDRG LVWPNGLTLDYEQNLIYWADANLEKIERMDLEHYLREVIVSRANSPFGLAIYGQYVYWTD LLTQKIYRANKVDGSGQTAVTVTLPFRPKGIRAVVKDQQQCLSPCDRFNGGCSHICAPGP NGAECQCPHEGRWYLANNNKYCIQDNGTRCDSSKFTCLSGKCIPDQLQCNDIDDCGDSSD ELETLCAFHSCPSTSFTCANGRCVPYSDRCDHYNDCGDNSDEAGCHFRACNRTEFTCSNG RCIPSELVCDGVDNCLDNSASDEKNCPERTCHTGYVKCTNSTICIPRSFLCDGDNDCGDM SDENPLFCATRSCGSDEFHCTSGPCIPARWYCDHEKDCSDGSDEPPTCEFSQSTCASDYF KCDNNRCIPMMWVCDGDNDCGDMSDEDERHNCKNRNCSSSEFACEVGVRPHRGCIPKSWV CDGEADCLDALDEHQNCTRRSCFGTEFVCNNGLCIPNHFRCDRNNDCGDYSDERGCVYPT CDETLFTCQNGLCINKAYVCDGDNDCKDNSDELEHLCHTPETTCPPHQFRCDNGNCIEMM KVCNNFPDCSDNSDEKGCGINECNDPTLSGCNQNCTDTLTSFYCSCNPGYKLLSDKRTCV DIDECEETPFVCSQKCENLPGTYICKCAPGYIREPDGKTCRQNSNIEPDLIFSNRYYLRN LTVYGHLYSLILQGLGNVVALDFDRVEKRLYWLDIENKVIERMFVNTTNRETVLKYNLPG AESLAVDWVTRKLYWVDSYLNCLSVSDLNGRYRRKLAEHCVDANNTFCFENPRGLALHPR YGHVYWADWGDRAYIGRVGMDGTLKSLIISTKIMWPNGLTIDYTNDLLYWADAHLGYIEF SDLEGRHRHTVYETGTLSHPFAITIFEDTIYWTDWNTKTVEKGNKYNGSDRVALLNVTHR PYDIRVYHPYRQPIVPNPCGTNNGGCSHLCLIKEGGVGFTCECPDNFYTVQRGPNTQCLP MCSSTQFLCADSEMCIPIWWKCDGRRDCLDGSDEPITCPQRFCALGMFQCNDGNCTNSHS LCNLRQDCPDGSDEDPVLCEHHQCEPYEWQCANKRCIPESWQCDMQDDCDDNSDEDSSHC ASRTCRPGYFKCANGHCIPQIWKCDVDNDCGDYSDEPLQECLGPSYRCDNYTEFSCKTNY RCIPKWAVCNGVDDCRDNSDEQDCESMTCKPSGEFRCTNHRCIPLRWRCDGQNDCGDRSD EENCAPRKCTESEFRCDDQSCIPSRWVCDQTNDCGDNSDERDCEMMTCRPGYFQCDSGHC ISEHMKCNGVADCRDASDEANCPTRFPNGAYCPATMFECKNHVCIHSSWKCDGDNDCGDG SDEELHLCLNVACDSPYRFRCDNNRCIYRHEVCNQEDDCGDGSDEKKELCVEPTPRPCTP DEFKCSNGRCIPQHRVCDHVNDCGDNFDETGCNTGKDRSCAENLCEHNCTQLREGGFICS CRPGFKPNSIDRNLCEDINECMQFGSCPQICHNTKGSYECSCAEGFTSLSDRYGERCAAD GSPPLLLLPENVRIRKYNLSSLQFSEYLEDQERIKAMDYDWDPEGTGLSVVYYTVLGEGS NSGAIKRAYIPNFESGSNNPVMEINLDLKYIVQPDGLAVDWVGRHIYWSDARRQRIEVAE LDGRYRKWLISTELGQPAAIVVNPKLGFMYWTDWGENPKIESAWMDGQRRKVLVQEDLGW PTGLCIDYMNGDRIYWSDLKDNIVETIKYDGTDRRIVVTSAVNPYSLDIFESQLYWTSKD KGEVWIQDKFGRNKKEKLLTVNPWLTQVRVFHQRKYNHSVPNRCKDVCSHLCLLRPKGYT CACPQGSRFLEGSVTVCDAAIVGAVSMPPPCRCMNRGSCYFDENNLPKCKCSSGYVGEYC EMGLSQGVPPGTTASVLLTVILIVIIAALATLGFLHYRKTGSILPSLPKLSSLSHLKSSE NGNGVTFRSGDDVNMDIGVTGFGPESAIDRSLAMNEHFAADFGKSPIIFENPTYSSKDTA ITVAQPTTAPVTESATVYNKNYGSPINPAELVTDTKPTSSSDETQPTKWNIFKRKPKQNT NFENPFYSEMENEPKVGAAVTPPPSPSPPAKVSWKKGSTPGYSATEDTFKDTANLVREDS EA LRP226935120 c.2363G>C mutation site aa.788 missense mutation: CRSNEFLCQPGLCITASWRCDGTRDCPNGADEIDCPTSSCHSNQFLCPNEQLCIPESWVC DGEDDCNDGADERRHCPGITCSSRQFTCENGECIPGEFRCDHSTDCLDGTDEKNCRYPVC EQLTCADGACYNTSQRCNGQVDCRDASDERNCTHRCTRTEFQCGSGQCIPRTYVCDHEID CEDGSDEHSCPAYRTCKGNEFTCPNGVCIAQSWVCDGESDCVDNADEDGCESKINRTFEC YPNEWACPKSGKCIPISKVCDGTLDCPGGEDESNITAGQQCDVNLCPSLGCEYQCHRSPG GGMCYCPSGFIVNQNRTNNCVDFDDCQIWGVCDQLCEDRIGHHRCNCVEGYVLERQKHCR ANSSFGEAFVIFSNGRNLLKGSIRGNNFQILAESQNRGLAVGVDFHYRLRRVFWTDIVQK KVFSVDISGSRIREVLGVSIEDPENLAVDWVNNKLYIVETRVNCIDVVDLDGSHRITLIS EYLGHPRGIAVDPTVGYLFFSDWQNVFGVPRIERAYMDGSNRKDLVTTKLGWPGGITLDL VSKRVYWVDSRFDYIETVTYDGLKRMTVIHGGADIPHPFSISLFEGQLFFTDWTKMAVLK ANKFKETNPRLYYRSSLKPFGVTVYHGLRQPYARNPCAHENGGCQHICVLSHRTFNGGLG YRCKCRLGYIPDWDDYHCVAAERFLLFSSNLAVRGIPLTLSHQTEVILPVTGSSSIFLGI DFDAREKAIFFSDTKKNIIYRQKLDGTGREIITANRVPAVQSLSFDWISRNLYWTDSSYR SVSVIRLADKSRRTIIQNLNNPQSIVVHPTAGYIFFSVWYRPAKILRAWADGSNILPIVN TTLGWPSGLSIDWSSSRLYWVDAFFDKIEHSNLDGSDRKALTNVHQLTHPFGLAVSQDYI YVTDWRRGIIRFGKYNPGQSIILRSGVGSVMRAKVYDSRVQTGSNACSRPTNPNGDCSHF CFPVPNSQRVCGCPYGMSLASDHLTCVENASREPPVEQCGTLSFSCHNGRCVPLQYRCDG FDDCLDNSDEAQCTTSNATCSPLAFECKREGHCIPSMWRCDGEDDCLDGSDEQNCPTRAP TSCRADQFTCDNNFCIPRSWVCDTDNDCKDGSDEKSCNYTQTCSPTQFHCPDHRCIALTF VCDGTKDCADGSDEIGCVINCTASQFTCVSNGQCISKTYRCDGVFDCDDHSDETDCPTRP PGMCHQDEFQCQEDGICIPKTWECDGHEDCLQGSDEHNGCPPKTCHPSHFVCQNGNCIYR NWLCDGDNDCGDMSDEKDCPTQPFQCPSWQWQCPGHSICVNLSAVCDGVSDCPGGTDESP LCNQNSCSDSNGGCTHQCIQGPYGAQCQCPLGYLLGNDSKTCEDIDECRTPGFCSQYCYN MRGSFRCWCDSEYTLDADRRTCKATGNEKSETLLLVASQSQLVAGNMTQNGYFVYPVIQH GSHIVAVDFDSVSGRIFWSDGTQGKIWSAFQNGTDKKLILDSGVSMTGSIAVDWIGRNLY WTDISLRTIDVAKLDGSHKTVLISENITNLGGLAVDPRATDRVMFWSDWGSHPRIERASM DGSQRTIIVQEKIYWPNGLALDYPNRLLYFMDGYLDYLDFCHYDGSNRRQVIASDLILRH PYSISLFEDTVYWSDRATHEVMKANKWHGGNQSVVMTLHQPLGIVVVHPAKQPVSSNPCS YTRCSHLCLLSSKSLYSCACPSGWTLARDSVTCVRDDQAFLIVVRNSIIFGISLNPDVKT FDGMVPISGIRNGYDVAVDYSEQFIYWLENPGEIHRVKTDGTNRTVFAPLSSLGASASLA LDWLSRNLYFTDHVTRSIKVMTLQGDVSYRKTLIANDGTSLGVGLPVGITIDPINGKLYW SDRGTNSGLPPKIASANMDGKSPRTLFTGSLENVAFITLDIEEQKLYWAVSSTGVIERGN VDGTNRMILVNHLSYPWGLAVHGRFLYYSDEEYEVIERVDKATGANKVVLRNNLPDLRGL KIYQRRSESSNGCSNNMNACQQICLPVPGRLFSCACATGFKLNPDHQTCSPYNSFIVVST LRTIRGFSLQLSDHSEAMVPVAGPGRNALHVDVDVSSGFIYWCDFNISVASNNAIRRIKP DGSNFTNIVTDGIGVNGVRGIAVDWVAGNLYFTNAFRSETLIEVLRINTTHRHILLKTIV DMPRDIVVDPKNRYLFWSDYGQNPKIERSFLDCTNRTVLVSDITATPRGLALDHSSNYIY WVDDAVDLIARISIEGGETEVIRFGSHYPAPYAITVFGNSIIWVDRNLKKILQASKEPNR ADRPTVIRDNIDWLRDVTIFDQSVQPRSPAEVNNNPCLENNGGCAQFCFALPKSQTPKCD CAFGTLQADGKSCAISSENFLIFALDDSLRSLRFDPKDYSQPFPAISVERMAVALDYDSI DNRIYFTQLLPSGKGQISYINLNSRSSPPTVVVSGIGSPEGIAFDWINKRIYYSDYTNQM IKSIATDGSRHTLIAQVPKPRGIVLDPCQGYMYWTDWGTNAKIEMATMGGYSRRSLVDRG LVWPNGLTLDYEQNLIYWADANLEKIERMDLEHYLREVIVSRANSPFGLAIYGQYVYWTD LLTQKIYRANKVDGSGQTAVTVTLPFRPKGIRAVVKDQQQCLSPCDRFNGGCSHICAPGP NGAECQCPHEGRWYLANNNKYCIQDNGTRCDSSKFTCLSGKCIPDQLQCNDIDDCGDSSD ELETLCAFHSCPSTSFTCANGRCVPYSDRCDHYNDCGDNSDEAGCHFRACNRTEFTCSNG RCIPSELVCDGVDNCLDNSASDEKNCPERTCHTGYVKCTNSTICIPRSFLCDGDNDCGDM SDENPLFCATRSCGSDEFHCTSGPCIPARWYCDHEKDCSDGSDEPPTCEFSQSTCASDYF KCDNNRCIPMMWVCDGDNDCGDMSDEDERHNCKNRNCSSSEFACEVGVRPHRGCIPKSWV CDGEADCLDALDEHQNCTRRSCFGTEFVCNNGLCIPNHFRCDRNNDCGDYSDERGCVYPT CDETLFTCQNGLCINKAYVCDGDNDCKDNSDELEHLCHTPETTCPPHQFRCDNGNCIEMM KVCNNFPDCSDNSDEKGCGINECNDPTLSGCNQNCTDTLTSFYCSCNPGYKLLSDKRTCV DIDECEETPFVCSQKCENLPGTYICKCAPGYIREPDGKTCRQNSNIEPDLIFSNRYYLRN LTVYGHLYSLILQGLGNVVALDFDRVEKRLYWLDIENKVIERMFVNTTNRETVLKYNLPG AESLAVDWVTRKLYWVDSYLNCLSVSDLNGRYRRKLAEHCVDANNTFCFENPRGLALHPR YGHVYWADWGDRAYIGRVGMDGTLKSLIISTKIMWPNGLTIDYTNDLLYWADAHLGYIEF SDLEGRHRHTVYETGTLSHPFAITIFEDTIYWTDWNTKTVEKGNKYNGSDRVALLNVTHR PYDIRVYHPYRQPIVPNPCGTNNGGCSHLCLIKEGGVGFTCECPDNFYTVQRGPNTQCLP MCSSTQFLCADSEMCIPIWWKCDGRRDCLDGSDEPITCPQRFCALGMFQCNDGNCTNSHS LCNLRQDCPDGSDEDPVLCEHHQCEPYEWQCANKRCIPESWQCDMQDDCDDNSDEDSSHC ASRTCRPGYFKCANGHCIPQIWKCDVDNDCGDYSDEPLQECLGPSYRCDNYTEFSCKTNY RCIPKWAVCNGVDDCRDNSDEQDCESMTCKPSGEFRCTNHRCIPLRWRCDGQNDCGDRSD EENCAPRKCTESEFRCDDQSCIPSRWVCDQTNDCGDNSDERDCEMMTCRPGYFQCDSGHC ISEHMKCNGVADCRDASDEANCPTRFPNGAYCPATMFECKNHVCIHSSWKCDGDNDCGDG SDEELHLCLNVACDSPYRFRCDNNRCIYRHEVCNQEDDCGDGSDEKKELCVEPTPRPCTP DEFKCSNGRCIPQHRVCDHVNDCGDNFDETGCNTGKDRSCAENLCEHNCTQLREGGFICS CRPGFKPNSIDRNLCEDINECMQFGSCPQICHNTKGSYECSCAEGFTSLSDRYGERCAAD GSPPLLLLPENVRIRKYNLSSLQFSEYLEDQERIKAMDYDWDPEGTGLSVVYYTVLGEGS NSGAIKRAYIPNFESGSNNPVMEINLDLKYIVQPDGLAVDWVGRHIYWSDARRQRIEVAE LDGRYRKWLISTELGQPAAIVVNPKLGFMYWTDWGENPKIESAWMDGQRRKVLVQEDLGW PTGLCIDYMNGDRIYWSDLKDNIVETIKYDGTDRRIVVTSAVNPYSLDIFESQLYWTSKD KGEVWIQDKFGRNKKEKLLTVNPWLTQVRVFHQRKYNHSVPNRCKDVCSHLCLLRPKGYT CACPQGSRFLEGSVTVCDAAIVGAVSMPPPCRCMNRGSCYFDENNLPKCKCSSGYVGEYC EMGLSQGVPPGTTASVLLTVILIVIIAALATLGFLHYRKTGSILPSLPKLSSLSHLKSSE NGNGVTFRSGDDVNMDIGVTGFGPESAIDRSLAMNEHFAADFGKSPIIFENPTYSSKDTA ITVAQPTTAPVTESATVYNKNYGSPINPAELVTDTKPTSSSDETQPTKWNIFKRKPKQNT NFENPFYSEMENEPKVGAAVTPPPSPSPPAKVSWKKGSTPGYSATEDTFKDTANLVREDS EA LRP226937805 c.2617C>T mutation site aa.873 missense mutation: CRSNEFLCQPGLCITASWRCDGTRDCPNGADEIDCPTSSCHSNQFLCPNEQLCIPESWVC DGEDDCNDGADERRHCPGITCSSRQFTCENGECIPGEFRCDHSTDCLDGTDEKNCRYPVC EQLTCADGACYNTSQRCNGQVDCRDASDERNCTHRCTRTEFQCGSGQCIPRTYVCDHEID CEDGSDEHSCPAYRTCKGNEFTCPNGVCIAQSWVCDGESDCVDNADEDGCESKINRTFEC YPNEWACPKSGKCIPISKVCDGTLDCPGGEDESNITAGQQCDVNLCPSLGCEYQCHRSPG GGMCYCPSGFIVNQNRTNNCVDFDDCQIWGVCDQLCEDRIGHHRCNCVEGYVLERQKHCR ANSSFGEAFVIFSNGRNLLKGSIRGNNFQILAESQNRGLAVGVDFHYRLRRVFWTDIVQK KVFSVDISGSRIREVLGVSIEDPENLAVDWVNNKLYIVETRVNCIDVVDLDGSHRITLIS EYLGHPRGIAVDPTVGYLFFSDWQNVFGVPRIERAYMDGSNRKDLVTTKLGWPGGITLDL VSKRVYWVDSRFDYIETVTYDGLKRMTVIHGGADIPHPFSISLFEGQLFFTDWTKMAVLK ANKFKETNPRLYYRSSLKPFGVTVYHGLRQPYARNPCAHENGGCQHICVLSHRTFNGGLG YRCKCRLGYIPDWDDYHCVAAERFLLFSSNLAVRGIPLTLSHQTEVILPVTGSSSIFLGI DFDAREKAIFFSDTKKNIIYRQKLDGTGREIITANRVPAVQSLSFDWISRNLYWTDSSYR SVSVIRLGDKSRRTIIQNLNNPQSIVVHPTAGYIFFSVWYRPAKILRAWADGSNILPIVN TTLGWPSGLSIDWSSSRLYWVDAFFDKIEHSNFDGSDRKALTNVHQLTHPFGLAVSQDYI YVTDWRRGIIRFGKYNPGQSIILRSGVGSVMRAKVYDSRVQTGSNACSRPTNPNGDCSHF CFPVPNSQRVCGCPYGMSLASDHLTCVENASREPPVEQCGTLSFSCHNGRCVPLQYRCDG FDDCLDNSDEAQCTTSNATCSPLAFECKREGHCIPSMWRCDGEDDCLDGSDEQNCPTRAP TSCRADQFTCDNNFCIPRSWVCDTDNDCKDGSDEKSCNYTQTCSPTQFHCPDHRCIALTF VCDGTKDCADGSDEIGCVINCTASQFTCVSNGQCISKTYRCDGVFDCDDHSDETDCPTRP PGMCHQDEFQCQEDGICIPKTWECDGHEDCLQGSDEHNGCPPKTCHPSHFVCQNGNCIYR NWLCDGDNDCGDMSDEKDCPTQPFQCPSWQWQCPGHSICVNLSAVCDGVSDCPGGTDESP LCNQNSCSDSNGGCTHQCIQGPYGAQCQCPLGYLLGNDSKTCEDIDECRTPGFCSQYCYN MRGSFRCWCDSEYTLDADRRTCKATGNEKSETLLLVASQSQLVAGNMTQNGYFVYPVIQH GSHIVAVDFDSVSGRIFWSDGTQGKIWSAFQNGTDKKLILDSGVSMTGSIAVDWIGRNLY WTDISLRTIDVAKLDGSHKTVLISENITNLGGLAVDPRATDRVMFWSDWGSHPRIERASM DGSQRTIIVQEKIYWPNGLALDYPNRLLYFMDGYLDYLDFCHYDGSNRRQVIASDLILRH PYSISLFEDTVYWSDRATHEVMKANKWHGGNQSVVMTLHQPLGIVVVHPAKQPVSSNPCS YTRCSHLCLLSSKSLYSCACPSGWTLARDSVTCVRDDQAFLIVVRNSIIFGISLNPDVKT FDGMVPISGIRNGYDVAVDYSEQFIYWLENPGEIHRVKTDGTNRTVFAPLSSLGASASLA LDWLSRNLYFTDHVTRSIKVMTLQGDVSYRKTLIANDGTSLGVGLPVGITIDPINGKLYW SDRGTNSGLPPKIASANMDGKSPRTLFTGSLENVAFITLDIEEQKLYWAVSSTGVIERGN VDGTNRMILVNHLSYPWGLAVHGRFLYYSDEEYEVIERVDKATGANKVVLRNNLPDLRGL KIYQRRSESSNGCSNNMNACQQICLPVPGRLFSCACATGFKLNPDHQTCSPYNSFIVVST LRTIRGFSLQLSDHSEAMVPVAGPGRNALHVDVDVSSGFIYWCDFNISVASNNAIRRIKP DGSNFTNIVTDGIGVNGVRGIAVDWVAGNLYFTNAFRSETLIEVLRINTTHRHILLKTIV DMPRDIVVDPKNRYLFWSDYGQNPKIERSFLDCTNRTVLVSDITATPRGLALDHSSNYIY WVDDAVDLIARISIEGGETEVIRFGSHYPAPYAITVFGNSIIWVDRNLKKILQASKEPNR ADRPTVIRDNIDWLRDVTIFDQSVQPRSPAEVNNNPCLENNGGCAQFCFALPKSQTPKCD CAFGTLQADGKSCAISSENFLIFALDDSLRSLRFDPKDYSQPFPAISVERMAVALDYDSI DNRIYFTQLLPSGKGQISYINLNSRSSPPTVVVSGIGSPEGIAFDWINKRIYYSDYTNQM IKSIATDGSRHTLIAQVPKPRGIVLDPCQGYMYWTDWGTNAKIEMATMGGYSRRSLVDRG LVWPNGLTLDYEQNLIYWADANLEKIERMDLEHYLREVIVSRANSPFGLAIYGQYVYWTD LLTQKIYRANKVDGSGQTAVTVTLPFRPKGIRAVVKDQQQCLSPCDRFNGGCSHICAPGP NGAECQCPHEGRWYLANNNKYCIQDNGTRCDSSKFTCLSGKCIPDQLQCNDIDDCGDSSD ELETLCAFHSCPSTSFTCANGRCVPYSDRCDHYNDCGDNSDEAGCHFRACNRTEFTCSNG RCIPSELVCDGVDNCLDNSASDEKNCPERTCHTGYVKCTNSTICIPRSFLCDGDNDCGDM SDENPLFCATRSCGSDEFHCTSGPCIPARWYCDHEKDCSDGSDEPPTCEFSQSTCASDYF KCDNNRCIPMMWVCDGDNDCGDMSDEDERHNCKNRNCSSSEFACEVGVRPHRGCIPKSWV CDGEADCLDALDEHQNCTRRSCFGTEFVCNNGLCIPNHFRCDRNNDCGDYSDERGCVYPT CDETLFTCQNGLCINKAYVCDGDNDCKDNSDELEHLCHTPETTCPPHQFRCDNGNCIEMM KVCNNFPDCSDNSDEKGCGINECNDPTLSGCNQNCTDTLTSFYCSCNPGYKLLSDKRTCV DIDECEETPFVCSQKCENLPGTYICKCAPGYIREPDGKTCRQNSNIEPDLIFSNRYYLRN LTVYGHLYSLILQGLGNVVALDFDRVEKRLYWLDIENKVIERMFVNTTNRETVLKYNLPG AESLAVDWVTRKLYWVDSYLNCLSVSDLNGRYRRKLAEHCVDANNTFCFENPRGLALHPR YGHVYWADWGDRAYIGRVGMDGTLKSLIISTKIMWPNGLTIDYTNDLLYWADAHLGYIEF SDLEGRHRHTVYETGTLSHPFAITIFEDTIYWTDWNTKTVEKGNKYNGSDRVALLNVTHR PYDIRVYHPYRQPIVPNPCGTNNGGCSHLCLIKEGGVGFTCECPDNFYTVQRGPNTQCLP MCSSTQFLCADSEMCIPIWWKCDGRRDCLDGSDEPITCPQRFCALGMFQCNDGNCTNSHS LCNLRQDCPDGSDEDPVLCEHHQCEPYEWQCANKRCIPESWQCDMQDDCDDNSDEDSSHC ASRTCRPGYFKCANGHCIPQIWKCDVDNDCGDYSDEPLQECLGPSYRCDNYTEFSCKTNY RCIPKWAVCNGVDDCRDNSDEQDCESMTCKPSGEFRCTNHRCIPLRWRCDGQNDCGDRSD EENCAPRKCTESEFRCDDQSCIPSRWVCDQTNDCGDNSDERDCEMMTCRPGYFQCDSGHC ISEHMKCNGVADCRDASDEANCPTRFPNGAYCPATMFECKNHVCIHSSWKCDGDNDCGDG SDEELHLCLNVACDSPYRFRCDNNRCIYRHEVCNQEDDCGDGSDEKKELCVEPTPRPCTP DEFKCSNGRCIPQHRVCDHVNDCGDNFDETGCNTGKDRSCAENLCEHNCTQLREGGFICS CRPGFKPNSIDRNLCEDINECMQFGSCPQICHNTKGSYECSCAEGFTSLSDRYGERCAAD GSPPLLLLPENVRIRKYNLSSLQFSEYLEDQERIKAMDYDWDPEGTGLSVVYYTVLGEGS NSGAIKRAYIPNFESGSNNPVMEINLDLKYIVQPDGLAVDWVGRHIYWSDARRQRIEVAE LDGRYRKWLISTELGQPAAIVVNPKLGFMYWTDWGENPKIESAWMDGQRRKVLVQEDLGW PTGLCIDYMNGDRIYWSDLKDNIVETIKYDGTDRRIVVTSAVNPYSLDIFESQLYWTSKD KGEVWIQDKFGRNKKEKLLTVNPWLTQVRVFHQRKYNHSVPNRCKDVCSHLCLLRPKGYT CACPQGSRFLEGSVTVCDAAIVGAVSMPPPCRCMNRGSCYFDENNLPKCKCSSGYVGEYC EMGLSQGVPPGTTASVLLTVILIVIIAALATLGFLHYRKTGSILPSLPKLSSLSHLKSSE NGNGVTFRSGDDVNMDIGVTGFGPESAIDRSLAMNEHFAADFGKSPIIFENPTYSSKDTA ITVAQPTTAPVTESATVYNKNYGSPINPAELVTDTKPTSSSDETQPTKWNIFKRKPKQNT NFENPFYSEMENEPKVGAAVTPPPSPSPPAKVSWKKGSTPGYSATEDTFKDTANLVREDS EA LRP226963305c.5471A>G mutation site aa.1824 missense mutation: CRSNEFLCQPGLCITASWRCDGTRDCPNGADEIDCPTSSCHSNQFLCPNEQLCIPESWVC DGEDDCNDGADERRHCPGITCSSRQFTCENGECIPGEFRCDHSTDCLDGTDEKNCRYPVC EQLTCADGACYNTSQRCNGQVDCRDASDERNCTHRCTRTEFQCGSGQCIPRTYVCDHEID CEDGSDEHSCPAYRTCKGNEFTCPNGVCIAQSWVCDGESDCVDNADEDGCESKINRTFEC YPNEWACPKSGKCIPISKVCDGTLDCPGGEDESNITAGQQCDVNLCPSLGCEYQCHRSPG GGMCYCPSGFIVNQNRTNNCVDFDDCQIWGVCDQLCEDRIGHHRCNCVEGYVLERQKHCR ANSSFGEAFVIFSNGRNLLKGSIRGNNFQILAESQNRGLAVGVDFHYRLRRVFWTDIVQK KVFSVDISGSRIREVLGVSIEDPENLAVDWVNNKLYIVETRVNCIDVVDLDGSHRITLIS EYLGHPRGIAVDPTVGYLFFSDWQNVFGVPRIERAYMDGSNRKDLVTTKLGWPGGITLDL VSKRVYWVDSRFDYIETVTYDGLKRMTVIHGGADIPHPFSISLFEGQLFFTDWTKMAVLK ANKFKETNPRLYYRSSLKPFGVTVYHGLRQPYARNPCAHENGGCQHICVLSHRTFNGGLG YRCKCRLGYIPDWDDYHCVAAERFLLFSSNLAVRGIPLTLSHQTEVILPVTGSSSIFLGI DFDAREKAIFFSDTKKNIIYRQKLDGTGREIITANRVPAVQSLSFDWISRNLYWTDSSYR SVSVIRLGDKSRRTIIQNLNNPQSIVVHPTAGYIFFSVWYRPAKILRAWADGSNILPIVN TTLGWPSGLSIDWSSSRLYWVDAFFDKIEHSNLDGSDRKALTNVHQLTHPFGLAVSQDYI YVTDWRRGIIRFGKYNPGQSIILRSGVGSVMRAKVYDSRVQTGSNACSRPTNPNGDCSHF CFPVPNSQRVCGCPYGMSLASDHLTCVENASREPPVEQCGTLSFSCHNGRCVPLQYRCDG FDDCLDNSDEAQCTTSNATCSPLAFECKREGHCIPSMWRCDGEDDCLDGSDEQNCPTRAP TSCRADQFTCDNNFCIPRSWVCDTDNDCKDGSDEKSCNYTQTCSPTQFHCPDHRCIALTF VCDGTKDCADGSDEIGCVINCTASQFTCVSNGQCISKTYRCDGVFDCDDHSDETDCPTRP PGMCHQDEFQCQEDGICIPKTWECDGHEDCLQGSDEHNGCPPKTCHPSHFVCQNGNCIYR NWLCDGDNDCGDMSDEKDCPTQPFQCPSWQWQCPGHSICVNLSAVCDGVSDCPGGTDESP LCNQNSCSDSNGGCTHQCIQGPYGAQCQCPLGYLLGNDSKTCEDIDECRTPGFCSQYCYN MRGSFRCWCDSEYTLDADRRTCKATGNEKSETLLLVASQSQLVAGNMTQNGYFVYPVIQH GSHIVAVDFDSVSGRIFWSDGTQGKIWSAFQNGTDKKLILDSGVSMTGSIAVDWIGRNLY WTDISLRTIDVAKLDGSHKTVLISENITNLGGLAVDPRATDRVMFWSDWGSHPRIERASM DGSQRTIIVQEKIYWPNGLALDYPNRLLYFMDGYLDYLDFCHYDGSNRRQVIASDLILRH PYSISLFEDTVYWSDRATHEVMKANKWHGGNQSVVMTLHQPLGIVVVHPAKQPVSSNPCS YTRCSHLCLLSSKSLYSCACPSGWTLARDSVTCVRDDQAFLIVVRNSIIFGISLNPDVKT FDGMVPISGIRNGYDVAVDYSEQFIYWLENPGEIHRVKTDGTNRTVFAPLSSLGASASLA LDWLSRNLYFTDHVTRSIKVMTLRGDVSYRKTLIANDGTSLGVGLPVGITIDPINGKLYW SDRGTNSGLPPKIASANMDGKSPRTLFTGSLENVAFITLDIEEQKLYWAVSSTGVIERGN VDGTNRMILVNHLSYPWGLAVHGRFLYYSDEEYEVIERVDKATGANKVVLRNNLPDLRGL KIYQRRSESSNGCSNNMNACQQICLPVPGRLFSCACATGFKLNPDHQTCSPYNSFIVVST LRTIRGFSLQLSDHSEAMVPVAGPGRNALHVDVDVSSGFIYWCDFNISVASNNAIRRIKP DGSNFTNIVTDGIGVNGVRGIAVDWVAGNLYFTNAFRSETLIEVLRINTTHRHILLKTIV DMPRDIVVDPKNRYLFWSDYGQNPKIERSFLDCTNRTVLVSDITATPRGLALDHSSNYIY WVDDAVDLIARISIEGGETEVIRFGSHYPAPYAITVFGNSIIWVDRNLKKILQASKEPNR ADRPTVIRDNIDWLRDVTIFDQSVQPRSPAEVNNNPCLENNGGCAQFCFALPKSQTPKCD CAFGTLQADGKSCAISSENFLIFALDDSLRSLRFDPKDYSQPFPAISVERMAVALDYDSI DNRIYFTQLLPSGKGQISYINLNSRSSPPTVVVSGIGSPEGIAFDWINKRIYYSDYTNQM IKSIATDGSRHTLIAQVPKPRGIVLDPCQGYMYWTDWGTNAKIEMATMGGYSRRSLVDRG LVWPNGLTLDYEQNLIYWADANLEKIERMDLEHYLREVIVSRANSPFGLAIYGQYVYWTD LLTQKIYRANKVDGSGQTAVTVTLPFRPKGIRAVVKDQQQCLSPCDRFNGGCSHICAPGP NGAECQCPHEGRWYLANNNKYCIQDNGTRCDSSKFTCLSGKCIPDQLQCNDIDDCGDSSD ELETLCAFHSCPSTSFTCANGRCVPYSDRCDHYNDCGDNSDEAGCHFRACNRTEFTCSNG RCIPSELVCDGVDNCLDNSASDEKNCPERTCHTGYVKCTNSTICIPRSFLCDGDNDCGDM SDENPLFCATRSCGSDEFHCTSGPCIPARWYCDHEKDCSDGSDEPPTCEFSQSTCASDYF KCDNNRCIPMMWVCDGDNDCGDMSDEDERHNCKNRNCSSSEFACEVGVRPHRGCIPKSWV CDGEADCLDALDEHQNCTRRSCFGTEFVCNNGLCIPNHFRCDRNNDCGDYSDERGCVYPT CDETLFTCQNGLCINKAYVCDGDNDCKDNSDELEHLCHTPETTCPPHQFRCDNGNCIEMM KVCNNFPDCSDNSDEKGCGINECNDPTLSGCNQNCTDTLTSFYCSCNPGYKLLSDKRTCV DIDECEETPFVCSQKCENLPGTYICKCAPGYIREPDGKTCRQNSNIEPDLIFSNRYYLRN LTVYGHLYSLILQGLGNVVALDFDRVEKRLYWLDIENKVIERMFVNTTNRETVLKYNLPG AESLAVDWVTRKLYWVDSYLNCLSVSDLNGRYRRKLAEHCVDANNTFCFENPRGLALHPR YGHVYWADWGDRAYIGRVGMDGTLKSLIISTKIMWPNGLTIDYTNDLLYWADAHLGYIEF SDLEGRHRHTVYETGTLSHPFAITIFEDTIYWTDWNTKTVEKGNKYNGSDRVALLNVTHR PYDIRVYHPYRQPIVPNPCGTNNGGCSHLCLIKEGGVGFTCECPDNFYTVQRGPNTQCLP MCSSTQFLCADSEMCIPIWWKCDGRRDCLDGSDEPITCPQRFCALGMFQCNDGNCTNSHS LCNLRQDCPDGSDEDPVLCEHHQCEPYEWQCANKRCIPESWQCDMQDDCDDNSDEDSSHC ASRTCRPGYFKCANGHCIPQIWKCDVDNDCGDYSDEPLQECLGPSYRCDNYTEFSCKTNY RCIPKWAVCNGVDDCRDNSDEQDCESMTCKPSGEFRCTNHRCIPLRWRCDGQNDCGDRSD EENCAPRKCTESEFRCDDQSCIPSRWVCDQTNDCGDNSDERDCEMMTCRPGYFQCDSGHC ISEHMKCNGVADCRDASDEANCPTRFPNGAYCPATMFECKNHVCIHSSWKCDGDNDCGDG SDEELHLCLNVACDSPYRFRCDNNRCIYRHEVCNQEDDCGDGSDEKKELCVEPTPRPCTP DEFKCSNGRCIPQHRVCDHVNDCGDNFDETGCNTGKDRSCAENLCEHNCTQLREGGFICS CRPGFKPNSIDRNLCEDINECMQFGSCPQICHNTKGSYECSCAEGFTSLSDRYGERCAAD GSPPLLLLPENVRIRKYNLSSLQFSEYLEDQERIKAMDYDWDPEGTGLSVVYYTVLGEGS NSGAIKRAYIPNFESGSNNPVMEINLDLKYIVQPDGLAVDWVGRHIYWSDARRQRIEVAE LDGRYRKWLISTELGQPAAIVVNPKLGFMYWTDWGENPKIESAWMDGQRRKVLVQEDLGW PTGLCIDYMNGDRIYWSDLKDNIVETIKYDGTDRRIVVTSAVNPYSLDIFESQLYWTSKD KGEVWIQDKFGRNKKEKLLTVNPWLTQVRVFHQRKYNHSVPNRCKDVCSHLCLLRPKGYT CACPQGSRFLEGSVTVCDAAIVGAVSMPPPCRCMNRGSCYFDENNLPKCKCSSGYVGEYC EMGLSQGVPPGTTASVLLTVILIVIIAALATLGFLHYRKTGSILPSLPKLSSLSHLKSSE NGNGVTFRSGDDVNMDIGVTGFGPESAIDRSLAMNEHFAADFGKSPIIFENPTYSSKDTA ITVAQPTTAPVTESATVYNKNYGSPINPAELVTDTKPTSSSDETQPTKWNIFKRKPKQNT NFENPFYSEMENEPKVGAAVTPPPSPSPPAKVSWKKGSTPGYSATEDTFKDTANLVREDS EA LRP226970787 c.6259A>G mutation site aa.2087 missense mutation: CRSNEFLCQPGLCITASWRCDGTRDCPNGADEIDCPTSSCHSNQFLCPNEQLCIPESWVC DGEDDCNDGADERRHCPGITCSSRQFTCENGECIPGEFRCDHSTDCLDGTDEKNCRYPVC EQLTCADGACYNTSQRCNGQVDCRDASDERNCTHRCTRTEFQCGSGQCIPRTYVCDHEID CEDGSDEHSCPAYRTCKGNEFTCPNGVCIAQSWVCDGESDCVDNADEDGCESKINRTFEC YPNEWACPKSGKCIPISKVCDGTLDCPGGEDESNITAGQQCDVNLCPSLGCEYQCHRSPG GGMCYCPSGFIVNQNRTNNCVDFDDCQIWGVCDQLCEDRIGHHRCNCVEGYVLERQKHCR ANSSFGEAFVIFSNGRNLLKGSIRGNNFQILAESQNRGLAVGVDFHYRLRRVFWTDIVQK KVFSVDISGSRIREVLGVSIEDPENLAVDWVNNKLYIVETRVNCIDVVDLDGSHRITLIS EYLGHPRGIAVDPTVGYLFFSDWQNVFGVPRIERAYMDGSNRKDLVTTKLGWPGGITLDL VSKRVYWVDSRFDYIETVTYDGLKRMTVIHGGADIPHPFSISLFEGQLFFTDWTKMAVLK ANKFKETNPRLYYRSSLKPFGVTVYHGLRQPYARNPCAHENGGCQHICVLSHRTFNGGLG YRCKCRLGYIPDWDDYHCVAAERFLLFSSNLAVRGIPLTLSHQTEVILPVTGSSSIFLGI DFDAREKAIFFSDTKKNIIYRQKLDGTGREIITANRVPAVQSLSFDWISRNLYWTDSSYR SVSVIRLGDKSRRTIIQNLNNPQSIVVHPTAGYIFFSVWYRPAKILRAWADGSNILPIVN TTLGWPSGLSIDWSSSRLYWVDAFFDKIEHSNLDGSDRKALTNVHQLTHPFGLAVSQDYI YVTDWRRGIIRFGKYNPGQSIILRSGVGSVMRAKVYDSRVQTGSNACSRPTNPNGDCSHF CFPVPNSQRVCGCPYGMSLASDHLTCVENASREPPVEQCGTLSFSCHNGRCVPLQYRCDG FDDCLDNSDEAQCTTSNATCSPLAFECKREGHCIPSMWRCDGEDDCLDGSDEQNCPTRAP TSCRADQFTCDNNFCIPRSWVCDTDNDCKDGSDEKSCNYTQTCSPTQFHCPDHRCIALTF VCDGTKDCADGSDEIGCVINCTASQFTCVSNGQCISKTYRCDGVFDCDDHSDETDCPTRP PGMCHQDEFQCQEDGICIPKTWECDGHEDCLQGSDEHNGCPPKTCHPSHFVCQNGNCIYR NWLCDGDNDCGDMSDEKDCPTQPFQCPSWQWQCPGHSICVNLSAVCDGVSDCPGGTDESP LCNQNSCSDSNGGCTHQCIQGPYGAQCQCPLGYLLGNDSKTCEDIDECRTPGFCSQYCYN MRGSFRCWCDSEYTLDADRRTCKATGNEKSETLLLVASQSQLVAGNMTQNGYFVYPVIQH GSHIVAVDFDSVSGRIFWSDGTQGKIWSAFQNGTDKKLILDSGVSMTGSIAVDWIGRNLY WTDISLRTIDVAKLDGSHKTVLISENITNLGGLAVDPRATDRVMFWSDWGSHPRIERASM DGSQRTIIVQEKIYWPNGLALDYPNRLLYFMDGYLDYLDFCHYDGSNRRQVIASDLILRH PYSISLFEDTVYWSDRATHEVMKANKWHGGNQSVVMTLHQPLGIVVVHPAKQPVSSNPCS YTRCSHLCLLSSKSLYSCACPSGWTLARDSVTCVRDDQAFLIVVRNSIIFGISLNPDVKT FDGMVPISGIRNGYDVAVDYSEQFIYWLENPGEIHRVKTDGTNRTVFAPLSSLGASASLA LDWLSRNLYFTDHVTRSIKVMTLQGDVSYRKTLIANDGTSLGVGLPVGITIDPINGKLYW SDRGTNSGLPPKIASANMDGKSPRTLFTGSLENVAFITLDIEEQKLYWAVSSTGVIERGN VDGTNRMILVNHLSYPWGLAVHGRFLYYSDEEYEVIERVDKATGANKVVLRNNLPDLRGL KIYQRRSESSNGCSNNMNACQQICLPVPGRLFSCACATGFKLNPDHQTCSPYNSFIVVST LRTIRGFSLQLSDHSEAMVPVAGPGRNALHVDVDVSSGFIYWCDFNVSVASNNAIRRIKP DGSNFTNIVTDGIGVNGVRGIAVDWVAGNLYFTNAFRSETLIEVLRINTTHRHILLKTIV DMPRDIVVDPKNRYLFWSDYGQNPKIERSFLDCTNRTVLVSDITATPRGLALDHSSNYIY WVDDAVDLIARISIEGGETEVIRFGSHYPAPYAITVFGNSIIWVDRNLKKILQASKEPNR ADRPTVIRDNIDWLRDVTIFDQSVQPRSPAEVNNNPCLENNGGCAQFCFALPKSQTPKCD CAFGTLQADGKSCAISSENFLIFALDDSLRSLRFDPKDYSQPFPAISVERMAVALDYDSI DNRIYFTQLLPSGKGQISYINLNSRSSPPTVVVSGIGSPEGIAFDWINKRIYYSDYTNQM IKSIATDGSRHTLIAQVPKPRGIVLDPCQGYMYWTDWGTNAKIEMATMGGYSRRSLVDRG LVWPNGLTLDYEQNLIYWADANLEKIERMDLEHYLREVIVSRANSPFGLAIYGQYVYWTD LLTQKIYRANKVDGSGQTAVTVTLPFRPKGIRAVVKDQQQCLSPCDRFNGGCSHICAPGP NGAECQCPHEGRWYLANNNKYCIQDNGTRCDSSKFTCLSGKCIPDQLQCNDIDDCGDSSD ELETLCAFHSCPSTSFTCANGRCVPYSDRCDHYNDCGDNSDEAGCHFRACNRTEFTCSNG RCIPSELVCDGVDNCLDNSASDEKNCPERTCHTGYVKCTNSTICIPRSFLCDGDNDCGDM SDENPLFCATRSCGSDEFHCTSGPCIPARWYCDHEKDCSDGSDEPPTCEFSQSTCASDYF KCDNNRCIPMMWVCDGDNDCGDMSDEDERHNCKNRNCSSSEFACEVGVRPHRGCIPKSWV CDGEADCLDALDEHQNCTRRSCFGTEFVCNNGLCIPNHFRCDRNNDCGDYSDERGCVYPT CDETLFTCQNGLCINKAYVCDGDNDCKDNSDELEHLCHTPETTCPPHQFRCDNGNCIEMM KVCNNFPDCSDNSDEKGCGINECNDPTLSGCNQNCTDTLTSFYCSCNPGYKLLSDKRTCV DIDECEETPFVCSQKCENLPGTYICKCAPGYIREPDGKTCRQNSNIEPDLIFSNRYYLRN LTVYGHLYSLILQGLGNVVALDFDRVEKRLYWLDIENKVIERMFVNTTNRETVLKYNLPG AESLAVDWVTRKLYWVDSYLNCLSVSDLNGRYRRKLAEHCVDANNTFCFENPRGLALHPR YGHVYWADWGDRAYIGRVGMDGTLKSLIISTKIMWPNGLTIDYTNDLLYWADAHLGYIEF SDLEGRHRHTVYETGTLSHPFAITIFEDTIYWTDWNTKTVEKGNKYNGSDRVALLNVTHR PYDIRVYHPYRQPIVPNPCGTNNGGCSHLCLIKEGGVGFTCECPDNFYTVQRGPNTQCLP MCSSTQFLCADSEMCIPIWWKCDGRRDCLDGSDEPITCPQRFCALGMFQCNDGNCTNSHS LCNLRQDCPDGSDEDPVLCEHHQCEPYEWQCANKRCIPESWQCDMQDDCDDNSDEDSSHC ASRTCRPGYFKCANGHCIPQIWKCDVDNDCGDYSDEPLQECLGPSYRCDNYTEFSCKTNY RCIPKWAVCNGVDDCRDNSDEQDCESMTCKPSGEFRCTNHRCIPLRWRCDGQNDCGDRSD EENCAPRKCTESEFRCDDQSCIPSRWVCDQTNDCGDNSDERDCEMMTCRPGYFQCDSGHC ISEHMKCNGVADCRDASDEANCPTRFPNGAYCPATMFECKNHVCIHSSWKCDGDNDCGDG SDEELHLCLNVACDSPYRFRCDNNRCIYRHEVCNQEDDCGDGSDEKKELCVEPTPRPCTP DEFKCSNGRCIPQHRVCDHVNDCGDNFDETGCNTGKDRSCAENLCEHNCTQLREGGFICS CRPGFKPNSIDRNLCEDINECMQFGSCPQICHNTKGSYECSCAEGFTSLSDRYGERCAAD GSPPLLLLPENVRIRKYNLSSLQFSEYLEDQERIKAMDYDWDPEGTGLSVVYYTVLGEGS NSGAIKRAYIPNFESGSNNPVMEINLDLKYIVQPDGLAVDWVGRHIYWSDARRQRIEVAE LDGRYRKWLISTELGQPAAIVVNPKLGFMYWTDWGENPKIESAWMDGQRRKVLVQEDLGW PTGLCIDYMNGDRIYWSDLKDNIVETIKYDGTDRRIVVTSAVNPYSLDIFESQLYWTSKD KGEVWIQDKFGRNKKEKLLTVNPWLTQVRVFHQRKYNHSVPNRCKDVCSHLCLLRPKGYT CACPQGSRFLEGSVTVCDAAIVGAVSMPPPCRCMNRGSCYFDENNLPKCKCSSGYVGEYC EMGLSQGVPPGTTASVLLTVILIVIIAALATLGFLHYRKTGSILPSLPKLSSLSHLKSSE NGNGVTFRSGDDVNMDIGVTGFGPESAIDRSLAMNEHFAADFGKSPIIFENPTYSSKDTA ITVAQPTTAPVTESATVYNKNYGSPINPAELVTDTKPTSSSDETQPTKWNIFKRKPKQNT NFENPFYSEMENEPKVGAAVTPPPSPSPPAKVSWKKGSTPGYSATEDTFKDTANLVREDS EA LRP226980105 c.8377A>C mutation site aa.2793 missense mutation: CRSNEFLCQPGLCITASWRCDGTRDCPNGADEIDCPTSSCHSNQFLCPNEQLCIPESWVC DGEDDCNDGADERRHCPGITCSSRQFTCENGECIPGEFRCDHSTDCLDGTDEKNCRYPVC EQLTCADGACYNTSQRCNGQVDCRDASDERNCTHRCTRTEFQCGSGQCIPRTYVCDHEID CEDGSDEHSCPAYRTCKGNEFTCPNGVCIAQSWVCDGESDCVDNADEDGCESKINRTFEC YPNEWACPKSGKCIPISKVCDGTLDCPGGEDESNITAGQQCDVNLCPSLGCEYQCHRSPG GGMCYCPSGFIVNQNRTNNCVDFDDCQIWGVCDQLCEDRIGHHRCNCVEGYVLERQKHCR ANSSFGEAFVIFSNGRNLLKGSIRGNNFQILAESQNRGLAVGVDFHYRLRRVFWTDIVQK KVFSVDISGSRIREVLGVSIEDPENLAVDWVNNKLYIVETRVNCIDVVDLDGSHRITLIS EYLGHPRGIAVDPTVGYLFFSDWQNVFGVPRIERAYMDGSNRKDLVTTKLGWPGGITLDL VSKRVYWVDSRFDYIETVTYDGLKRMTVIHGGADIPHPFSISLFEGQLFFTDWTKMAVLK ANKFKETNPRLYYRSSLKPFGVTVYHGLRQPYARNPCAHENGGCQHICVLSHRTFNGGLG YRCKCRLGYIPDWDDYHCVAAERFLLFSSNLAVRGIPLTLSHQTEVILPVTGSSSIFLGI DFDAREKAIFFSDTKKNIIYRQKLDGTGREIITANRVPAVQSLSFDWISRNLYWTDSSYR SVSVIRLGDKSRRTIIQNLNNPQSIVVHPTAGYIFFSVWYRPAKILRAWADGSNILPIVN TTLGWPSGLSIDWSSSRLYWVDAFFDKIEHSNLDGSDRKALTNVHQLTHPFGLAVSQDYI YVTDWRRGIIRFGKYNPGQSIILRSGVGSVMRAKVYDSRVQTGSNACSRPTNPNGDCSHF CFPVPNSQRVCGCPYGMSLASDHLTCVENASREPPVEQCGTLSFSCHNGRCVPLQYRCDG FDDCLDNSDEAQCTTSNATCSPLAFECKREGHCIPSMWRCDGEDDCLDGSDEQNCPTRAP TSCRADQFTCDNNFCIPRSWVCDTDNDCKDGSDEKSCNYTQTCSPTQFHCPDHRCIALTF VCDGTKDCADGSDEIGCVINCTASQFTCVSNGQCISKTYRCDGVFDCDDHSDETDCPTRP PGMCHQDEFQCQEDGICIPKTWECDGHEDCLQGSDEHNGCPPKTCHPSHFVCQNGNCIYR NWLCDGDNDCGDMSDEKDCPTQPFQCPSWQWQCPGHSICVNLSAVCDGVSDCPGGTDESP LCNQNSCSDSNGGCTHQCIQGPYGAQCQCPLGYLLGNDSKTCEDIDECRTPGFCSQYCYN MRGSFRCWCDSEYTLDARRTCKATGNEKSETLLLVASQSQLVAGNMTQNGYFVYPVIQH GSHIVAVDFDSVSGRIFWSDGTTQGKIWSAFQNGTDKKLILDSGVSMTGSIAVDWIGRNLY WTDISLRTIDVAKLDGSHKTVLISENITNLGGLAVDPRATDRVMFWSDWGSHPRIERASM DGSQRTIIVQEKIYWPNGLALDYPNRLLYFMDGYLDYLDFCHYDGSNRRQVIASDLILRH PYSISLFEDTVYWSDRATHEVMKANKWHGGNQSVVMTLHQPLGIVVVHPAKQPVSSNPCS YTRCSHLCLLSSKSLYSCACPSGWTLARDSVTCVRDDQAFLIVVRNSIIFGISLNPDVKT FDGMVPISGIRNGYDVAVDYSEQFIYWLENPGEIHRVKTDGTNRTVFAPLSSLGASASLA LDWLSRNLYFTDHHVTRSIKVMTLQGDVSYRKTLIANDGTSLGVGLPVGITIDPINGKLYW SDRGTNSGLPPKIASANMDGKSPRTLFTGSLENVAFITLDIEQKLYWAVSSTGVIERGN VDGTNRMILVNHLSYPWGLAVHGRFLYYSDEEYEVIERVDKATGANKVVLRNNLPDLRGL KIYQRRSESSNGCSNNMNACQQICLPVPGRLFSCACATGFKLNPDHQTCSPYNSFIVVST LRTIRGFSLQLSDHSEAMVPVAGPGRNALHVDVDVSSGFIYWCDFNISVASNNAIRRIKP DGSNFTNIVTDGIGVNGVRGIAVDWVAGNLYFTNAFRSETLIEVLRINTTHRHILLKTIV DMPRDIVVDPKNRYLFWSDYGQNPKIERSFLDCTNRTVLVSDITATPRGLALDHSSNYIY WVDDAVDLIARISIEGGETEVIRFGSHYPAPYAITVFGNSIIWVDRNLKKILQASKEPNR ADRPTVIRDNIDWLRDVTIFDQSVQPRSPAEVNNNPCLENNGGCAQFCFALPKSQTPKCD CAFGTLQADGKSCAISSENFLIFALDDSLRSLRFDPKDYSQPFPAISVERMAVALDYDSI DNRIYFTQLLPSGKGQISYINLNSRSSPPTVVVSGIGSPEGIAFDWINKRIYYSDYTNQM IKSIATDGSRHTLIAQVPKPRGIVLDPCQGYMYWTDWGTNAKIEMATMGGYSRRSLVDRG LVWPNGLTLDYEQNLIYWADANLEKIERMDLEHYLREVIVSRANSPFGLAIYGQYVYWTD LLTQKIYRANKVDGSGQTAVTVTLPFRPKGIRAVVKDQQQCLSPCDRFNGGCSHICAPGP NGAECQCPHEGRWYLANNNKYCIQDNGTRCDSSKFTCLSGKCIPDQLQCNDIDDCGDSSD ELETLCAFHSCPSTSFTCANGRCVPYSDRCDHYNDCGDNSDEAGCHFRACNRTEFTCSNG RCIPSELVCDGVDNCLDNSASDEKNCPERTCHPGYVKCTNSTICIPRSFLCDGDNDCGDM SDENPLFCATRSCGSDEFHCTSGPCIPARWYCDHEKDCSDGSDEPPTCEFSQSTCASDYF KCDNNRCIPMMWVCDGDNDCGDMSDEDERHNCKNRNCSSSEFACEVGVRPHRGCIPKSWV CDGEADCLDALDEHQNCTRRSCFGTEFVCNNGLCIPNHFRCDRNNDCGDYSDERGCVYPT CDETLFTCQNGLCINKAYVCDGDNDCKDNSDELEHLCHTPETTCPPHQFRCDNGNCIEMM KVCNNFPDCSDNSDEKGCGINECNDPTLSGCNQNCTDTLTSFYCSCNPGYKLLSDKRTCV DIDECEETPFVCSQKCENLPGTYICKCAPGYIREPDGKTCRQNSNIEPDLIFSNRYYLRN LTVYGHLYSLILQGLGNVVALDFDRVEKRLYWLDIENKVIERMFVNTTNRETVLKYNLPG AESLAVDWVTRKLYWVDSYLNCLSVSDLNGRYRRKLAEHCVDANNTFCFENPRGLALHPR YGHVYWADWGDRAYIGRVGMDGTLKSLIISTKIMWPNGLTIDYTNDLLYWADAHLGYIEF SDLEGRHRHTVYETGTLSHPFAITIFEDTIYWTDWNTKTVEKGNKYNGSDRVALLNVTHR PYDIRVYHPYRQPIVPNPCGTNNGGCSHLCLIKEGGVGFTCECPDNFYTVQRGPNTQCLP MCSSTQFLCADSEMCIPIWWKCDGRRDCLDGSDEPITCPQRFCALGMFQCNDGNCTNSHS LCNLRQDCPDGSDEDPVLCEHHQCEPYEWQCANKRCIPESWQCDMQDDCDDNSDEDSSHC ASRTCRPGYFKCANGHCIPQIWKCDVDNDCGDYSDEPLQECLGPSYRCDNYTEFSCKTNY RCIPKWAVCNGVDDCRDNSDEQDCESMTCKPSGEFRCTNHRCIPLRWRCDGQNDCGDRSD EENCAPRKCTESEFRCDDQSCIPSRWVCDQTNDCGDNSDERDCEMMTCRPGYFQCDSGHC ISEHMKCNGVADCRDASDEANCPTRFPNGAYCPATMFECKNHVCIHSSWKCDGDNDCGDG SDEELHLCLNVACDSPYRFRCDNNRCIYRHEVCNQEDDCGDGSDEKKELCVEPTPRPCTP DEFKCSNGRCIPQHRVCDHVNDCGDNFDETGCNTGKDRSCAENLCEHNCTQLREGGFICS CRPGFKPNSIDRNLCEDINECMQFGSCPQICHNTKGSYECSCAEGFTSLSDRYGERCAAD GSPPLLLLPENVRIRKYNLSSLQFSEYLEDQERIKAMDYDWDPEGTGLSVVYYTVLGEGS NSGAIKRAYIPNFESGSNNPVMEINLDLKYIVQPDGLAVDWVGRHIYWSDARRQRIEVAE LDGRYRKWLISTELGQPAAIVVNPKLGFMYWTDWGENPKIESAWMDGQRRKVLVQEDLGW PTGLCIDYMNGDRIYWSDLKDNIVETIKYDGTDRRIVVTSAVNPYSLDIFESQLYWTSKD KGEVWIQDKFGRNKKEKLLTVNPWLTQVRVFHQRKYNHSVPNRCKDVCSHLCLLRPKGYT CACPQGSRFLEGSVTVCDAAIVGAVSMPPPCRCMNRGSCYFDENNLPKCKCSSGYVGEYC EMGLSQGVPPGTTASVLLTVILIVIIAALATLGFLHYRKTGSILPSLPKLSSLSHLKSSE NGNGVTFRSGDDVNMDIGVTGFGPESAIDRSLAMNEHFAADFGKSPIIFENPTYSSKDTA ITVAQPTTAPVTESATVYNKNYGSPINPAELVTDTKPTSSSDETQPTKWNIFKRKPKQNT NFENPFYSEMENEPKVGAAVTPPPSPSPPAKVSWKKGSTPGYSATEDTFKDTANLVREDS EA LRP226988109 c.9706T>C mutation site aa.3236 missense mutation: CRSNEFLCQPGLCITASWRCDGTRDCPNGADEIDCPTSSCHSNQFLCPNEQLCIPESWVC DGEDDCNDGADERRHCPGITCSSRQFTCENGECIPGEFRCDHSTDCLDGTDEKNCRYPVC EQLTCADGACYNTSQRCNGQVDCRDASDERNCTHRCTRTEFQCGSGQCIPRTYVCDHEID CEDGSDEHSCPAYRTCKGNEFTCPNGVCIAQSWVCDGESDCVDNADEDGCESKINRTFEC YPNEWACPKSGKCIPISKVCDGTLDCPGGEDESNITAGQQCDVNLCPSLGCEYQCHRSPG GGMCYCPSGFIVNQNRTNNCVDFDDCQIWGVCDQLCEDRIGHHRCNCVEGYVLERQKHCR ANSSFGEAFVIFSNGRNLLKGSIRGNNFQILAESQNRGLAVGVDFHYRLRRVFWTDIVQK KVFSVDISGSRIREVLGVSIEDPENLAVDWVNNKLYIVETRVNCIDVVDLDGSHRITLIS EYLGHPRGIAVDPTVGYLFFSDWQNVFGVPRIERAYMDGSNRKDLVTTKLGWPGGITLDL VSKRVYWVDSRFDYIETVTYDGLKRMTVIHGGADIPHPFSISLFEGQLFFTDWTKMAVLK ANKFKETNPRLYYRSSLKPFGVTVYHGLRQPYARNPCAHENGGCQHICVLSHRTFNGGLG YRCKCRLGYIPDWDDYHCVAAERFLLFSSNLAVRGIPLTLSHQTEVILPVTGSSSIFLGI DFDAREKAIFFSDTKKNIIYRQKLDGTGREIITANRVPAVQSLSFDWISRNLYWTDSSYR SVSVIRLGDKSRRTIIQNLNNPQSIVVHPTAGYIFFSVWYRPAKILRAWADGSNILPIVN TTLGWPSGLSIDWSSSRLYWVDAFFDKIEHSNLDGSDRKALTNVHQLTHPFGLAVSQDYI YVTDWRRGIIRFGKYNPGQSIILRSGVGSVMRAKVYDSRVQTGSNACSRPTNPNGDCSHF CFPVPNSQRVCGCPYGMSLASDHLTCVENASREPPVEQCGTLSFSCHNGRCVPLQYRCDG FDDCLDNSDEAQCTTSNATCSPLAFECKREGHCIPSMWRCDGEDDCLDGSDEQNCPTRAP TSCRADQFTCDNNFCIPRSWVCDTDNDCKDGSDEKSCNYTQTCSPTQFHCPDHRCIALTF VCDGTKDCADGSDEIGCVINCTASQFTCVSNGQCISKTYRCDGVFDCDDHSDETDCPTRP PGMCHQDEFQCQEDGICIPKTWECDGHEDCLQGSDEHNGCPPKTCHPSHFVCQNGNCIYR NWLCDGDNDCGDMSDEKDCPTQPFQCPSWQWQCPGHSICVNLSAVCDGVSDCPGGTDESP LCNQNSCSDSNGGCTHQCIQGPYGAQCQCPLGYLLGNDSKTCEDIDECRTPGFCSQYCYN MRGSFRCWCDSEYTLDADRRTCKATGNEKSETLLLVASQSQLVAGNMTQNGYFVYPVIQH GSHIVAVDFDSVSGRIFWSDGTQGKIWSAFQNGTDKKLILDSGVSMTGSIAVDWIGRNLY WTDISLRTIDVAKLDGSHKTVLISENITNLGGLAVDPRATDRVMFWSDWGSHPRIERASM DGSQRTIIVQEKIYWPNGLALDYPNRLLYFMDGYLDYLDFCHYDGSNRRQVIASDLILRH PYSISLFEDTVYWSDRATHEVMKANKWHGGNQSVVMTLHQPLGIVVVHPAKQPVSSNPCS YTRCSHLCLLSSKSLYSCACPSGWTLARDSVTCVRDDQAFLIVVRNSIIFGISLNPDVKT FDGMVPISGIRNGYDVAVDYSEQFIYWLENPGEIHRVKTDGTNRTVFAPLSSLGASASLA LDWLSRNLYFTDHVTRSIKVMTLQGDVSYRKTLIANDGTSLGVGLPVGITIDPINGKLYW SDRGTNSGLPPKIASANMDGKSPRTLFTGSLENVAFITLDIEEQKLYWAVSSTGVIERGN VDGTNRMILVNHLSYPWGLAVHGRFLYYSDEEYEVIERVDKATGANKVVLRNNLPDLRGL KIYQRRSESSNGCSNNMNACQQICLPVPGRLFSCACATGFKLNPDHQTCSPYNSFIVVST LRTIRGFSLQLSDHSEAMVPVAGPGRNALHVDVDVSSGFIYWCDFNISVASNNAIRRIKP DGSNFTNIVTDGIGVNGVRGIAVDWVAGNLYFTNAFRSETLIEVLRINTTHRHILLKTIV DMPRDIVVDPKNRYLFWSDYGQNPKIERSFLDCTNRTVLVSDITATPRGLALDHSSNYIY WVDDAVDLIARISIEGGETEVIRFGSHYPAPYAITVFGNSIIWVDRNLKKILQASKEPNR ADRPTVIRDNIDWLRDVTIFDQSVQPRSPAEVNNNPCLENNGGCAQFCFALPKSQTPKCD CAFGTLQADGKSCAISSENFLIFALDDSLRSLRFDPKDYSQPFPAISVERMAVALDYDSI DNRIYFTQLLPSGKGQISYINLNSRSSPPTVVVSGIGSPEGIAFDWINKRIYYSDYTNQM IKSIATDGSRHTLIAQVPKPRGIVLDPCQGYMYWTDWGTNAKIEMATMGGYSRRSLVDRG LVWPNGLTLDYEQNLIYWADANLEKIERMDLEHYLREVIVSRANSPFGLAIYGQYVYWTD LLTQKIYRANKVDGSGQTAVTVTLPFRPKGIRAVVKDQQQCLSPCDRFNGGCSHICAPGP NGAECQCPHEGRWYLANNNKYCIQDNGTRCDSSKFTCLSGKCIPDQLQCNDIDDCGDSSD ELETLCAFHSCPSTSFTCANGRCVPYSDRCDHYNDCGDNSDEAGCHFRACNRTEFTCSNG RCIPSELVCDGVDNCLDNSASDEKNCPERTCHTGYVKCTNSTICIPRSFLCDGDNDCGDM SDENPLFCATRSCGSDEFHCTSGPCIPARWYCDHEKDCSDGSDEPPTCEFSQSTCASDYF KCDNNRCIPMMWVCDGDNDCGDMSDEDERHNCKNRNCSSSEFACEVGVRPHRGCIPKSWV CDGEADCLDALDEHQNCTRRSCFGTEFVCNNGLCIPNHFRCDRNNDCGDYSDERGCVYPT CDETLFTCQNGLCINKAYVCDGDNDCKDNSDELEHLCHTPETTCPPHQFRCDNGNCIEMM KVCNNFPDCSDNSDEKGCGINECNDPTLSGCNQNCTDTLTSFYCSCNPGYKLLSDKRTCV DIDECEETPFVCSQKCENLPGTYICKCAPGYIREPDGKTCRQNSNIEPDLIFSNRYYLRN LTVYGHLYSLILQGLGNVVALDFDRVEKRLYWLDIENKVIERMFVNTTNRETVLKHNLPG AESLAVDWVTRKLYWVDSYLNCLSVSDLNGRYRRKLAEHCVDANNTFCFENPRGLALHPR YGHVYWADWGDRAYIGRVGMDGTLKSLIISTKIMWPNGLTIDYTNDLLYWADAHLGYIEF SDLEGRHRHTVYETGTLSHPFAITIFEDTIYWTDWNTKTVEKGNKYNGSDRVALLNVTHR PYDIRVYHPYRQPIVPNPCGTNNGGCSHLCLIKEGGVGFTCECPDNFYTVQRGPNTQCLP MCSSTQFLCADSEMCIPIWWKCDGRRDCLDGSDEPITCPQRFCALGMFQCNDGNCTNSHS LCNLRQDCPDGSDEDPVLCEHHQCEPYEWQCANKRCIPESWQCDMQDDCDDNSDEDSSHC ASRTCRPGYFKCANGHCIPQIWKCDVDNDCGDYSDEPLQECLGPSYRCDNYTEFSCKTNY RCIPKWAVCNGVDDCRDNSDEQDCESMTCKPSGEFRCTNHRCIPLRWRCDGQNDCGDRSD EENCAPRKCTESEFRCDDQSCIPSRWVCDQTNDCGDNSDERDCEMMTCRPGYFQCDSGHC ISEHMKCNGVADCRDASDEANCPTRFPNGAYCPATMFECKNHVCIHSSWKCDGDNDCGDG SDEELHLCLNVACDSPYRFRCDNNRCIYRHEVCNQEDDCGDGSDEKKELCVEPTPRPCTP DEFKCSNGRCIPQHRVCDHVNDCGDNFDETGCNTGKDRSCAENLCEHNCTQLREGGFICS CRPGFKPNSIDRNLCEDINECMQFGSCPQICHNTKGSYECSCAEGFTSLSDRYGERCAAD GSPPLLLLPENVRIRKYNLSSLQFSEYLEDQERIKAMDYDWDPEGTGLSVVYYTVLGEGS NSGAIKRAYIPNFESGSNNPVMEINLDLKYIVQPDGLAVDWVGRHIYWSDARRQRIEVAE LDGRYRKWLISTELGQPAAIVVNPKLGFMYWTDWGENPKIESAWMDGQRRKVLVQEDLGW PTGLCIDYMNGDRIYWSDLKDNIVETIKYDGTDRRIVVTSAVNPYSLDIFESQLYWTSKD KGEVWIQDKFGRNKKEKLLTVNPWLTQVRVFHQRKYNHSVPNRCKDVCSHLCLLRPKGYT CACPQGSRFLEGSVTVCDAAIVGAVSMPPPCRCMNRGSCYFDENNLPKCKCSSGYVGEYC EMGLSQGVPPGTTASVLLTVILIVIIAALATLGFLHYRKTGSILPSLPKLSSLSHLKSSE NGNGVTFRSGDDVNMDIGVTGFGPESAIDRSLAMNEHFAADFGKSPIIFENPTYSSKDTA ITVAQPTTAPVTESATVYNKNYGSPINPAELVTDTKPTSSSDETQPTKWNIFKRKPKQNT NFENPFYSEMENEPKVGAAVTPPPSPSPPAKVSWKKGSTPGYSATEDTFKDTANLVREDS EA LRP226993170 c.10079T>A mutation site aa.3360 missense mutation: CRSNEFLCQPGLCITASWRCDGTRDCPNGADEIDCPTSSCHSNQFLCPNEQLCIPESWVC DGEDDCNDGADERRHCPGITCSSRQFTCENGECIPGEFRCDHSTDCLDGTDEKNCRYPVC EQLTCADGACYNTSQRCNGQVDCRDASDERNCTHRCTRTEFQCGSGQCIPRTYVCDHEID CEDGSDEHSCPAYRTCKGNEFTCPNGVCIAQSWVCDGESDCVDNADEDGCESKINRTFEC YPNEWACPKSGKCIPISKVCDGTLDCPGGEDESNITAGQQCDVNLCPSLGCEYQCHRSPG GGMCYCPSGFIVNQNRTNNCVDFDDCQIWGVCDQLCEDRIGHHRCNCVEGYVLERQKHCR ANSSFGEAFVIFSNGRNLLKGSIRGNNFQILAESQNRGLAVGVDFHYRLRRVFWTDIVQK KVFSVDISGSRIREVLGVSIEDPENLAVDWVNNKLYIVETRVNCIDVVDLDGSHRITLIS EYLGHPRGIAVDPTVGYLFFSDWQNVFGVPRIERAYMDGSNRKDLVTTKLGWPGGITLDL VSKRVYWVDSRFDYIETVTYDGLKRMTVIHGGADIPHPFSISLFEGQLFFTDWTKMAVLK ANKFKETNPRLYYRSSLKPFGVTVYHGLRQPYARNPCAHENGGCQHICVLSHRTFNGGLG YRCKCRLGYIPDWDDYHCVAAERFLLFSSNLAVRGIPLTLSHQTEVILPVTGSSSIFLGI DFDAREKAIFFSDTKKNIIYRQKLDGTGREIITANRVPAVQSLSFDWISRNLYWTDSSYR SVSVIRLGDKSRRTIIQNLNNPQSIVVHPTAGYIFFSVWYRPAKILRAWADGSNILPIVN TTLGWPSGLSIDWSSSRLYWVDAFFDKIEHSNLDGSDRKALTNVHQLTHPFGLAVSQDYI YVTDWRRGIIRFGKYNPGQSIILRSGVGSVMRAKVYDSRVQTGSNACSRPTNPNGDCSHF CFPVPNSQRVCGCPYGMSLASDHLTCVENASREPPVEQCGTLSFSCHNGRCVPLQYRCDG FDDCLDNSDEAQCTTSNATCSPLAFECKREGHCIPSMWRCDGEDDCLDGSDEQNCPTRAP TSCRADQFTCDNNFCIPRSWVCDTDNDCKDGSDEKSCNYTQTCSPTQFHCPDHRCIALTF VCDGTKDCADGSDEIGCVINCTASQFTCVSNGQCISKTYRCDGVFDCDDHSDETDCPTRP PGMCHQDEFQCQEDGICIPKTWECDGHEDCLQGSDEHNGCPPKTCHPSHFVCQNGNCIYR NWLCDGDNDCGDMSDEKDCPTQPFQCPSWQWQCPGHSICVNLSAVCDGVSDCPGGTDESP LCNQNSCSDSNGGCTHQCIQGPYGAQCQCPLGYLLGNDSKTCEDIDECRTPGFCSQYCYN MRGSFRCWCDSEYTLDADRRTCKATGNEKSETLLLVASQSQLVAGNMTQNGYFVYPVIQH GSHIVAVDFDSVSGRIFWSDGTQGKIWSAFQNGTDKKLILDSGVSMTGSIAVDWIGRNLY WTDISLRTIDVAKLDGSHKTVLISENITNLGGLAVDPRATDRVMFWSDWGSHPRIERASM DGSQRTIIVQEKIYWPNGLALDYPNRLLYFMDGYLDYLDFCHYDGSNRRQVIASDLILRH PYSISLFEDTVYWSDRATHEVMKANKWHGGNQSVVMTLHQPLGIVVVHPAKQPVSSNPCS YTRCSHLCLLSSKSLYSCACPSGWTLARDSVTCVRDDQAFLIVVRNSIIFGISLNPDVKT FDGMVPISGIRNGYDVAVDYSEQFIYWLENPGEIHRVKTDGTNRTVFAPLSSLGASASLA LDWLSRNLYFTDHVTRSIKVMTLQGDVSYRKTLIANDGTSLGVGLPVGITIDPINGKLYW SDRGTNSGLPPKIASANMDGKSPRTLFTGSLENVAFITLDIEEQKLYWAVSSTGVIERGN VDGTNRMILVNHLSYPWGLAVHGRFLYYSDEEYEVIERVDKATGANKVVLRNNLPDLRGL KIYQRRSESSNGCSNNMNACQQICLPVPGRLFSCACATGFKLNPDHQTCSPYNSFIVVST LRTIRGFSLQLSDHSEAMVPVAGPGRNALHVDVDVSSGFIYWCDFNISVASNNAIRRIKP DGSNFTNIVTDGIGVNGVRGIAVDWVAGNLYFTNAFRSETLIEVLRINTTHRHILLKTIV DMPRDIVVDPKNRYLFWSDYGQNPKIERSFLDCTNRTVLVSDITATPRGLALDHSSNYIY WVDDAVDLIARISIEGGETEVIRFGSHYPAPYAITVFGNSIIWVDRNLKKILQASKEPNR ADRPTVIRDNIDWLRDVTIFDQSVQPRSPAEVNNNPCLENNGGCAQFCFALPKSQTPKCD CAFGTLQADGKSCAISSENFLIFALDDSLRSLRFDPKDYSQPFPAISVERMAVALDYDSI DNRIYFTQLLPSGKGQISYINLNSRSSPPTVVVSGIGSPEGIAFDWINKRIYYSDYTNQM IKSIATDGSRHTLIAQVPKPRGIVLDPCQGYMYWTDWGTNAKIEMATMGGYSRRSLVDRG LVWPNGLTLDYEQNLIYWADANLEKIERMDLEHYLREVIVSRANSPFGLAIYGQYVYWTD LLTQKIYRANKVDGSGQTAVTVTLPFRPKGIRAVVKDQQQCLSPCDRFNGGCSHICAPGP NGAECQCPHEGRWYLANNNKYCIQDNGTRCDSSKFTCLSGKCIPDQLQCNDIDDCGDSSD ELETLCAFHSCPSTSFTCANGRCVPYSDRCDHYNDCGDNSDEAGCHFRACNRTEFTCSNG RCIPSELVCDGVDNCLDNSASDEKNCPERTCHTGYVKCTNSTICIPRSFLCDGDNDCGDM SDENPLFCATRSCGSDEFHCTSGPCIPARWYCDHEKDCSDGSDEPPTCEFSQSTCASDYF KCDNNRCIPMMWVCDGDNDCGDMSDEDERHNCKNRNCSSSEFACEVGVRPHRGCIPKSWV CDGEADCLDALDEHQNCTRRSCFGTEFVCNNGLCIPNHFRCDRNNDCGDYSDERGCVYPT CDETLFTCQNGLCINKAYVCDGDNDCKDNSDELEHLCHTPETTCPPHQFRCDNGNCIEMM KVCNNFPDCSDNSDEKGCGINECNDPTLSGCNQNCTDTLTSFYCSCNPGYKLLSDKRTCV DIDECEETPFVCSQKCENLPGTYICKCAPGYIREPDGKTCRQNSNIEPDLIFSNRYYLRN LTVYGHLYSLILQGLGNVVALDFDRVEKRLYWLDIENKVIERMFVNTTNRETVLKYNLPG AESLAVDWVTRKLYWVDSYLNCLSVSDLNGRYRRKLAEHCVDANNTFCFENPRGLALHPR YGHVYWADWGDRAYIGRVGMDGTLKSLIISTKIMWPNGLTIDYTNDLLYWADAHLGYIEY SDLEGRHRHTVYETGTLSHPFAITIFEDTIYWTDWNTKTVEKGNKYNGSDRVALLNVTHR PYDIRVYHPYRQPIVPNPCGTNNGGCSHLCLIKEGGVGFTCECPDNFYTVQRGPNTQCLP MCSSTQFLCADSEMCIPIWWKCDGRRDCLDGSDEPITCPQRFCALGMFQCNDGNCTNSHS LCNLRQDCPDGSDEDPVLCEHHQCEPYEWQCANKRCIPESWQCDMQDDCDDNSDEDSSHC ASRTCRPGYFKCANGHCIPQIWKCDVDNDCGDYSDEPLQECLGPSYRCDNYTEFSCKTNY RCIPKWAVCNGVDDCRDNSDEQDCESMTCKPSGEFRCTNHRCIPLRWRCDGQNDCGDRSD EENCAPRKCTESEFRCDDQSCIPSRWVCDQTNDCGDNSDERDCEMMTCRPGYFQCDSGHC ISEHMKCNGVADCRDASDEANCPTRFPNGAYCPATMFECKNHVCIHSSWKCDGDNDCGDG SDEELHLCLNVACDSPYRFRCDNNRCIYRHEVCNQEDDCGDGSDEKKELCVEPTPRPCTP DEFKCSNGRCIPQHRVCDHVNDCGDNFDETGCNTGKDRSCAENLCEHNCTQLREGGFICS CRPGFKPNSIDRNLCEDINECMQFGSCPQICHNTKGSYECSCAEGFTSLSDRYGERCAAD GSPPLLLLPENVRIRKYNLSSLQFSEYLEDQERIKAMDYDWDPEGTGLSVVYYTVLGEGS NSGAIKRAYIPNFESGSNNPVMEINLDLKYIVQPDGLAVDWVGRHIYWSDARRQRIEVAE LDGRYRKWLISTELGQPAAIVVNPKLGFMYWTDWGENPKIESAWMDGQRRKVLVQEDLGW PTGLCIDYMNGDRIYWSDLKDNIVETIKYDGTDRRIVVTSAVNPYSLDIFESQLYWTSKD KGEVWIQDKFGRNKKEKLLTVNPWLTQVRVFHQRKYNHSVPNRCKDVCSHLCLLRPKGYT CACPQGSRFLEGSVTVCDAAIVGAVSMPPPCRCMNRGSCYFDENNLPKCKCSSGYVGEYC EMGLSQGVPPGTTASVLLTVILIVIIAALATLGFLHYRKTGSILPSLPKLSSLSHLKSSE NGNGVTFRSGDDVNMDIGVTGFGPESAIDRSLAMNEHFAADFGKSPIIFENPTYSSKDTA ITVAQPTTAPVTESATVYNKNYGSPINPAELVTDTKPTSSSDETQPTKWNIFKRKPKQNT NFENPFYSEMENEPKVGAAVTPPPSPSPPAKVSWKKGSTPGYSATEDTFKDTANLVREDS EA LRP227001344c.11519G>A mutation site aa.3840 missense mutation: CRSNEFLCQPGLCITASWRCDGTRDCPNGADEIDCPTSSCHSNQFLCPNEQLCIPESWVC DGEDDCNDGADERRHCPGITCSSRQFTCENGECIPGEFRCDHSTDCLDGTDEKNCRYPVC EQLTCADGACYNTSQRCNGQVDCRDASDERNCTHRCTRTEFQCGSGQCIPRTYVCDHEID CEDGSDEHSCPAYRTCKGNEFTCPNGVCIAQSWVCDGESDCVDNADEDGCESKINRTFEC YPNEWACPKSGKCIPISKVCDGTLDCPGGEDESNITAGQQCDVNLCPSLGCEYQCHRSPG GGMCYCPSGFIVNQNRTNNCVDFDDCQIWGVCDQLCEDRIGHHRCNCVEGYVLERQKHCR ANSSFGEAFVIFSNGRNLLKGSIRGNNFQILAESQNRGLAVGVDFHYRLRRVFWTDIVQK KVFSVDISGSRIREVLGVSIEDPENLAVDWVNNKLYIVETRVNCIDVVDLDGSHRITLIS EYLGHPRGIAVDPTVGYLFFSDWQNVFGVPRIERAYMDGSNRKDLVTTKLGWPGGITLDL VSKRVYWVDSRFDYIETVTYDGLKRMTVIHGGADIPHPFSISLFEGQLFFTDWTKMAVLK ANKFKETNPRLYYRSSLKPFGVTVYHGLRQPYARNPCAHENGGCQHICVLSHRTFNGGLG YRCKCRLGYIPDWDDYHCVAAERFLLFSSNLAVRGIPLTLSHQTEVILPVTGSSSIFLGI DFDAREKAIFFSDTKKNIIYRQKLDGTGREIITANRVPAVQSLSFDWISRNLYWTDSSYR SVSVIRLGDKSRRTIIQNLNNPQSIVVHPTAGYIFFSVWYRPAKILRAWADGSNILPIVN TTLGWPSGLSIDWSSSRLYWVDAFFDKIEHSNLDGSDRKALTNVHQLTHPFGLAVSQDYI YVTDWRRGIIRFGKYNPGQSIILRSGVGSVMRAKVYDSRVQTGSNACSRPTNPNGDCSHF CFPVPNSQRVCGCPYGMSLASDHLTCVENASREPPVEQCGTLSFSCHNGRCVPLQYRCDG FDDCLDNSDEAQCTTSNATCSPLAFECKREGHCIPSMWRCDGEDDCLDGSDEQNCPTRAP TSCRADQFTCDNNFCIPRSWVCDTDNDCKDGSDEKSCNYTQTCSPTQFHCPDHRCIALTF VCDGTKDCADGSDEIGCVINCTASQFTCVSNGQCISKTYRCDGVFDCDDHSDETDCPTRP PGMCHQDEFQCQEDGICIPKTWECDGHEDCLQGSDEHNGCPPKTCHPSHFVCQNGNCIYR NWLCDGDNDCGDMSDEKDCPTQPFQCPSWQWQCPGHSICVNLSAVCDGVSDCPGGTDESP LCNQNSCSDSNGGCTHQCIQGPYGAQCQCPLGYLLGNDSKTCEDIDECRTPGFCSQYCYN MRGSFRCWCDSEYTLDADRRTCKATGNEKSETLLLVASQSQLVAGNMTQNGYFVYPVIQH GSHIVAVDFDSVSGRIFWSDGTQGKIWSAFQNGTDKKLILDSGVSMTGSIAVDWIGRNLY WTDISLRTIDVAKLDGSHKTVLISENITNLGGLAVDPRATDRVMFWSDWGSHPRIERASM DGSQRTIIVQEKIYWPNGLALDYPNRLLYFMDGYLDYLDFCHYDGSNRRQVIASDLILRH PYSISLFEDTVYWSDRATHEVMKANKWHGGNQSVVMTLHQPLGIVVVHPAKQPVSSNPCS YTRCSHLCLLSSKSLYSCACPSGWTLARDSVTCVRDDQAFLIVVRNSIIFGISLNPDVKT FDGMVPISGIRNGYDVAVDYSEQFIYWLENPGEIHRVKTDGTNRTVFAPLSSLGASASLA LDWLSRNLYFTDHVTRSIKVMTLQGDVSYRKTLIANDGTSLGVGLPVGITIDPINGKLYW SDRGTNSGLPPKIASANMDGKSPRTLFTGSLENVAFITLDIEEQKLYWAVSSTGVIERGN VDGTNRMILVNHLSYPWGLAVHGRFLYYSDEEYEVIERVDKATGANKVVLRNNLPDLRGL KIYQRRSESSNGCSNNMNACQQICLPVPGRLFSCACATGFKLNPDHQTCSPYNSFIVVST LRTIRGFSLQLSDHSEAMVPVAGPGRNALHVDVDVSSGFIYWCDFNISVASNNAIRRIKP DGSNFTNIVTDGIGVNGVRGIAVDWVAGNLYFTNAFRSETLIEVLRINTTHRHILLKTIV DMPRDIVVDPKNRYLFWSDYGQNPKIERSFLDCTNRTVLVSDITATPRGLALDHSSNYIY WVDDAVDLIARISIEGGETEVIRFGSHYPAPYAITVFGNSIIWVDRNLKKILQASKEPNR ADRPTVIRDNIDWLRDVTIFDQSVQPRSPAEVNNNPCLENNGGCAQFCFALPKSQTPKCD CAFGTLQADGKSCAISSENFLIFALDDSLRSLRFDPKDYSQPFPAISVERMAVALDYDSI DNRIYFTQLLPSGKGQISYINLNSRSSPPTVVVSGIGSPEGIAFDWINKRIYYSDYTNQM IKSIATDGSRHTLIAQVPKPRGIVLDPCQGYMYWTDWGTNAKIEMATMGGYSRRSLVDRG LVWPNGLTLDYEQNLIYWADANLEKIERMDLEHYLREVIVSRANSPFGLAIYGQYVYWTD LLTQKIYRANKVDGSGQTAVTVTLPFRPKGIRAVVKDQQQCLSPCDRFNGGCSHICAPGP NGAECQCPHEGRWYLANNNKYCIQDNGTRCDSSKFTCLSGKCIPDQLQCNDIDDCGDSSD ELETLCAFHSCPSTSFTCANGRCVPYSDRCDHYNDCGDNSDEAGCHFRACNRTEFTCSNG RCIPSELVCDGVDNCLDNSASDEKNCPERTCHTGYVKCTNSTICIPRSFLCDGDNDCGDM SDENPLFCATRSCGSDEFHCTSGPCIPARWYCDHEKDCSDGSDEPPTCEFSQSTCASDYF KCDNNRCIPMMWVCDGDNDCGDMSDEDERHNCKNRNCSSSEFACEVGVRPHRGCIPKSWV CDGEADCLDALDEHQNCTRRSCFGTEFVCNNGLCIPNHFRCDRNNDCGDYSDERGCVYPT CDETLFTCQNGLCINKAYVCDGDNDCKDNSDELEHLCHTPETTCPPHQFRCDNGNCIEMM KVCNNFPDCSDNSDEKGCGINECNDPTLSGCNQNCTDTLTSFYCSCNPGYKLLSDKRTCV DIDECEETPFVCSQKCENLPGTYICKCAPGYIREPDGKTCRQNSNIEPDLIFSNRYYLRN LTVYGHLYSLILQGLGNVVALDFDRVEKRLYWLDIENKVIERMFVNTTNRETVLKYNLPG AESLAVDWVTRKLYWVDSYLNCLSVSDLNGRYRRKLAEHCVDANNTFCFENPRGLALHPR YGHVYWADWGDRAYIGRVGMDGTLKSLIISTKIMWPNGLTIDYTNDLLYWADAHLGYIEF SDLEGRHRHTVYETGTLSHPFAITIFEDTIYWTDWNTKTVEKGNKYNGSDRVALLNVTHR PYDIRVYHPYRQPIVPNPCGTNNGGCSHLCLIKEGGVGFTCECPDNFYTVQRGPNTQCLP MCSSTQFLCADSEMCIPIWWKCDGRRDCLDGSDEPITCPQRFCALGMFQCNDGNCTNSHS LCNLRQDCPDGSDEDPVLCEHHQCEPYEWQCANKRCIPESWQCDMQDDCDDNSDEDSSHC ASRTCRPGYFKCANGHCIPQIWKCDVDNDCGDYSDEPLQECLGPSYRCDNYTEFSCKTNY RCIPKWAVCNGVDDCRDNSDEQDCESMTCKPSGEFRCTNHRCIPLRWRCDGQNDCGDRSD EENCAPRKCTESEFRCDDQSCIPSRWVCDQTNDCGDNSDERDCEMMTCRPGYFQCDSGHC ISEHMKCNGVADCRDASDEANCPTRFPNGAYCPATMFECKNHVCIHSSWKCDGDNDCGDD SDEELHLCLNVACDSPYRFRCDNNRCIYRHEVCNQEDDCGDGSDEKKELCVEPTPRPCTP DEFKCSNGRCIPQHRVCDHVNDCGDNFDETGCNTGKDRSCAENLCEHNCTQLREGGFICS CRPGFKPNSIDRNLCEDINECMQFGSCPQICHNTKGSYECSCAEGFTSLSDRYGERCAAD GSPPLLLLPENVRIRKYNLSSLQFSEYLEDQERIKAMDYDWDPEGTGLSVVYYTVLGEGS NSGAIKRAYIPNFESGSNNPVMEINLDLKYIVQPDGLAVDWVGRHIYWSDARRQRIEVAE LDGRYRKWLISTELGQPAAIVVNPKLGFMYWTDWGENPKIESAWMDGQRRKVLVQEDLGW PTGLCIDYMNGDRIYWSDLKDNIVETIKYDGTDRRIVVTSAVNPYSLDIFESQLYWTSKD KGEVWIQDKFGRNKKEKLLTVNPWLTQVRVFHQRKYNHSVPNRCKDVCSHLCLLRPKGYT CACPQGSRFLEGSVTVCDAAIVGAVSMPPPCRCMNRGSCYFDENNLPKCKCSSGYVGEYC EMGLSQGVPPGTTASVLLTVILIVIIAALATLGFLHYRKTGSILPSLPKLSSLSHLKSSE NGNGVTFRSGDDVNMDIGVTGFGPESAIDRSLAMNEHFAADFGKSPIIFENPTYSSKDTA ITVAQPTTAPVTESATVYNKNYGSPINPAELVTDTKPTSSSDETQPTKWNIFKRKPKQNT NFENPFYSEMENEPKVGAAVTPPPSPSPPAKVSWKKGSTPGYSATEDTFKDTANLVREDS EA The present invention and its embodiments have been described above. This description is not restrictive, and the embodiments shown throughout are only one of the embodiments of the present invention. The actual structure is not limited to this. In conclusion, if those skilled in the art are inspired by this description and design similar structures and embodiments without departing from the spirit of the present invention, they should all fall within the protection scope of the present invention.

Claims

1. A SNP molecular marker associated with the economic traits of Pinan cattle, characterized in that, The molecular marker is located in the exon region of the APLF gene on bovine chromosome 11, specifically at the rs135687714 site, where the base is mutated from T to C, causing a missense mutation of the amino acid from Met to Thr.

2. The SNP molecular marker related to the economic traits of Pinan cattle according to claim 1, characterized in that, The molecular marker is linked to the rs108964133 locus on chromosome 11 in the cattleQTL database (R²=0.5), and the two can be used as a combined molecular marker for Pinan cattle breeding.

3. The SNP molecular marker related to the economic traits of Pinan cattle according to claim 1, characterized in that, The PCR primer pair for the SNP molecular marker associated with the economic traits of Pinan cattle has the following primer sequences: SEQ ID NO.2 (forward primer): 5'-CAGGACTGCTTGAAGATGGA-3'; SEQ ID NO.3 (reverse primer): 5'-TGGTGTTCTCAGCCTCTTTC-3'.

4. The SNP molecular marker related to the economic traits of Pinan cattle according to claim 1, characterized in that, The aforementioned KASP primer pair for SNP molecular markers related to the economic traits of Pinan cattle includes: SEQ ID NO.4 (Allele T-specific primer, FAM tail): 5'-GAAGGTGACCAAGTTCATGCT–TCTCAGGACTGCTTGAAGATGGT-3'; SEQ ID NO.5 (allele C-specific primer, HEX tail): 5'-GAAGGTCGGAGTCAACGGATT–TCTCAGGACTGCTTGAAGATGGC-3'; SEQ ID NO.6 (Universal reverse primer): 5'-AGCAGGAAGGACACTGACAC-3'.

5. A molecular detection method for primer pairs, characterized in that, The molecular detection method can perform genotyping at the rs135687714 locus in individual Pinan cattle, which can be used for MAS-assisted breeding.

6. The application of an SNP molecular marker or detection method in early molecular-assisted selection of Pinan cattle to identify individuals associated with 11 growth traits, including hip width, height, and body structure.

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