Immune trait associated protein of pig, encoding gene and application of the same
A technology of related proteins and coding genes, applied in the direction of animal/human peptides, peptide sources, etc., can solve problems such as pigs that have not been reported
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Embodiment 1
[0040] Embodiment 1, the acquisition of porcine immune traits-related protein (LBP) and its full-length coding region and partial genome sequence (pig LBP gene)
[0041] 1. Acquisition of porcine immune trait-related protein (LBP) and its full-length coding region
[0042] Using human LBP gene cDNA (GenBank accession number: NM_006263) as an information probe, using the BLAST tool in NCBI to perform homologous sequence screening in the GenBank pig EST database, to obtain a series of ESTs with a homology of more than 80% (the fragment length is greater than 100bp), then use the seqman program in DNAstar software to perform EST splicing sequence, and design primers according to the splicing sequence. The primer sequences are as follows:
[0043] F1: 5'-CTGCATCGGACCTGACCCA-3' (sequence 4 in the sequence listing)
[0044] R1: 5'-TGCCCTTCAAGAAAGTACCAGC-3' (sequence 5 in the sequence listing)
[0045] Extract the total RNA from the liver tissue of a Tongcheng pig, reverse transcr...
Embodiment 2
[0051] Example 2, Establishment of PCR-RFLP Diagnostic Method for Pig LBP Gene and Correlation Analysis of the Gene Marker Characters
[0052] 1. Establishment of PCR-RFLP diagnostic method for pig LBP gene
[0053] The partial genome gene sequence of pig LBP obtained in step 2 of Example 1 shows that the 191st nucleotide at the 5' end of sequence 3 in the sequence listing is a polymorphic site, which is A or G. When the 191st nucleotide at the 5' end of sequence 3 in the sequence listing is A, the gene is assumed to be controlled by the A allele; when the 191st nucleotide at the 5' end of sequence 3 in the sequence listing is G , the gene is assumed to be controlled by the B allele, and these two alleles can form three genotypes: pure and AA, BB, and hybrid AB.
[0054] When the 191st nucleotide at the 5' end of sequence 3 in the sequence listing is G, there is one MspI restriction site (C↓CGG), so the part of the pig LBP obtained by amplifying step 2 of Example 1 Genomic g...
Embodiment 3
[0063] Embodiment 3, the distribution of pig LBP gene genotype in different pig breeds
[0064] Different kinds of pigs (Wuzhishan pigs (34) shown in Table 1, Laiwu pigs (32), Tongcheng pigs (41), Landrace pigs (26), Large White pigs (26) were tested according to the method of Example 2. ), respectively purchased from the Wuzhishan Miniature Pig Breeding Farm of the Beijing Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences; the Laiwu Black Pig Breeding Farm of Laiwu City, Shandong Province; to analyze
[0065] The results are shown in Table 3. According to the results of the genotype and gene frequency in Table 3, the B allele is dominant in the three domestic varieties of Wuzhishan, Tongcheng and Laiwu tested; while in the foreign varieties Changbai and Dabai The A allele is dominant.
[0066] Table 3 Distribution of LBP 329G / A mutation Msp I polymorphism in 5 pig breeds
[0067]
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