天祝白牦牛ANGPTL4基因的单核苷酸多态性研究

Single Nucleotide Polymorphisms Detection of ANGPTL4 Gene in Tianzhu White Yak

[目的]本研究旨在对牛ANGPTL4基因进行SNPs检测,并研究其与天祝白牦牛部分经济性状的相关性。[方法]以36头天祝白牦牛为研究对象,采集其血样并提取基因组DNA,设计ANGPTL4基因的特异引物对P1和P2,对ANGPTL4基因的部分序列进行扩增,并利用PCR-SSCP结合测序的方法检测扩增产物序列的多态性。[结果]以自行设计的上述两对引物扩增分别获得长为260 bp（位于GenBank公布序列NC-007305.2的296~555 bp间）和170 bp（位于序列NC-007305.2的4 610~4 779 bp间）的天祝白牦牛ANGPTL4基因的两个片段。引物对P1的扩增产物经变性聚丙烯酰胺凝胶电泳,出现两种带型：AA和AB。AA和AB两种基因型频率分别为0.9444和0.0556。多态信息含量是0.1000,为低度多态。对具有不同带型的个体测序结果表明,引物对P1的扩增片段上有两处发生碱基突变,分别位于ANGPTL4基因的第466 bp处（T→C）和479 bp处（T→C）,导致相应的氨基酸改变Cys→Arg,Phe→Ser。在引物对P2的扩增片段上没有发现SNPs位点。[结论]引物对P1的扩增位点可以尝试作为天祝白牦牛品质改良的辅助选择标记。

【Objective】The variation of ANGPTL4 gene was examined,and the association of this polymorphism with some economic traits was investigated in this study.【Method] 36 blood samples of TianZhu white yak were collected.The genomic DNA was extracted and two fragments of ANGPTL4gene were amplified with the use of designed primers（P1,P2） in this paper.And then,PCR-SSCP was conducted to detect the polymorphism of ANGPTL4 gene in TianZhu white yak herd.【Result】 Two fragments with the size of 260 bp（located at 296-555 bp of GenBank sequence numbered NC-007305.2）and 170 bp（located at 4610 bp-4779 bp of NC-007305.2） were obtained and two genotypes（AA and AB types）were detected in PCR product of primer pairs P1 after using the SDS polyacrylamide gel electrophoresis（PAGE）.Frequencies of genotype AA and AB was 0.9444 and 0.0556,respectively.The PIC value was 0.1000,which belonged to low degree polymorphism.Two single nucleotide substitutions,one was located at 466 bp（Y = C/T）（GenBank： NC-007305.2） and the other at 479 bp（Y = C/T） were detected in PCR product of P1 primer,resulting in amino acid changing with Cys→Arg,Phe→Ser.But no substitution was found in PCR product of P2 primer.【Conclusion】 We could conclude that the site of P1 primer could be a candidate molecular marker for the quality improvment of TianZhu white yak.