Cloning, sequencing and identification of single nucleotide polymorphisms of partial sequence on the porcine CACNA1S gene

文献类型： 外文期刊

第一作者： Fang XiaoMin

作者： Fang XiaoMin;Ren ShouWen;Fang XiaoMin;Xu NingYing

作者机构：

关键词： pig;calcium channel alpha(1) subunit (CACNA1S);single nucleotide polymorphism (SNP);PCR-SSCP;PCR-RFLP

期刊名称：SCIENCE IN CHINA SERIES C-LIFE SCIENCES （ 影响因子：1.61； 五年影响因子：1.148 ）

ISSN： 1006-9305

年卷期： 2008 年 51 卷 4 期

页码：

收录情况： SCI

摘要： CACNA1S gene encodes the alpha(1) subunit of the calcium channel. The mutation of CACNA1S gene can cause hypokalemic periodic paralysis (HypoKPP) and maliglant hyperthermia synarome (MHS) in human beings. Current research on CACNA1S was mainly in human being and model animal, but rarely in livestock and poultry. In this study, Yorkshire pigs (23), Pietrain pigs (30), Jinhua pigs (115) and the second generation (126) of crossbred of Jinhua and Pietrain were used. Primers were designed according to the sequence of human CACNA1S gene and PCR was carried out using pig genome DNA. PCR products were sequenced and compared with that of human, and then single nucleotide polymorphisms (SNPs) were investigated by PCR-SSCP, while PCR-RFLP tests were performed to validate the mutations. Results indicated: (1) the 5211 bp DNA fragments of porcine CACNA1S gene were acquired (GenBank accession number: DQ767693 ) and the identity of the exon region was 82.6% between human and pig; (2) fifty-seven mutations were found within the cloned sequences, among which 24 were in exon region; (3) the results of PCR-RFLP were in accordance with that of PCR-SSCP. According to the EST of porcine CACNA1S gene published in GenBank (Bx914582, Bx666997), 8 of the 11 SNPs identified in the present study were consistent with the base difference between two EST fragments.

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