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Molecular cloning and application of pig backfat thickness related SLC13A5 gene

A gene and base technology, applied in the field of nucleic acid molecules comprising pig gene nucleotides as shown in SEQ ID NO: 1

Active Publication Date: 2015-02-11
HUNAN CHUWEIXIANG AGRICUTLTURE CO LTD
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  • Abstract
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  • Application Information

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Problems solved by technology

[0006] At present, there is no report on the difference in the expression level of pig SLC13A5 gene between different pig species and different tissues at home and abroad, and there are few reports related to this gene. This invention uses RACE (Rapid Amplification cDNA Ends) technology to clone pig SLC13A5 gene for the first time The full length of the cDNA, and the SNP of the gene was screened, and the PCR-RFLP technique was used to type, and the correlation between the gene and the pig's corrected backfat thickness and the age of 100kg was analyzed

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  • Molecular cloning and application of pig backfat thickness related SLC13A5 gene

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Embodiment Construction

[0033] The present invention will be further explained below in conjunction with specific examples, but the specific implementation does not limit the present invention in any way.

[0034] Using the human SLC13A5 gene sequence (GenBank accession number NM_001284509.1) as the seed sequence, using BLASTN in GenBank, referring to pig EST (Expressed Sequence Tags) with more than 80% homology to design specific primers, using RACE technology to clone The full-length cDNA of the pig SLC13A5 gene was produced. The polymorphism of the SLC13A5 gene was designed by using comparative genomics methods based on the sequence of the human SLC13A5 gene. There is a base mutation of A / G, resulting in PCR-RFLP-BSu36I polymorphism, and using this to detect the distribution of the marker in Chinese and foreign pig breeds.

[0035] The longissimus dorsi muscle of 25-day-old Shaziling pigs was taken, and the total RNA was extracted and the first strand of cDNA was synthesized using SUPERSCRIPT II R...

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Abstract

The invention belongs to the technical field of livestock molecular biology, and particularly relates to molecular cloning and application of a pig backfat thickness related SLC13A5 gene. A molecular marker disclosed by the invention is obtained by SLC13A5 gene cloning, and a sequence marked by an SLC13A5 molecule is shown in SEQ ID NO:1. According to the polymorphism of the SLC13A5 gene, a primer is designed according to SLC13A5 gene sequence of human by using a comparative genomic method and amplified by taking pig genome DNA as a template; SNP of the amplified fragment is screened in a sequencing manner; PCR-RFLP is utilized to implement genetic typing; base mutation of one T / C at the 251bp is found, which causes PCR-RFLP-Bsu36I polymorphism; moreover, the marker is used for detecting the conditions of Chinese and foreign pig breeds. The invention provides new molecular marking for pig marker assisted selection.

Description

technical field [0001] The invention belongs to the technical field of livestock molecular biology, and relates to a nucleic acid molecule comprising porcine gene nucleotides as shown in SEQ ID NO:1. The present invention also relates to the molecular cloning method of the SLC13A5 gene shown in SEQ ID NO: 1, the single nucleotide polymorphism site in the polynucleotide sequence and the method for detecting the single nucleotide polymorphism site . Background technique [0002] Pork is the main source of animal protein. With the improvement of people's living standards, the requirements for meat quality also increase accordingly. There are genetic and non-genetic factors that affect pork quality. The fat content in pig muscle has a great influence on the meat quality. Generally speaking, the higher the fat content in the muscle, the tenderer the pork quality, and the better the meat quality. Most meat quality traits belong to quantitative traits, which are controlled by min...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 马海明王玲玉柴玉兰肖定福杨虎
Owner HUNAN CHUWEIXIANG AGRICUTLTURE CO LTD
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