山羊角蛋白14基因启动子分析及其多态性研究

Sequence Analysis and Polymorphism of K14 Gene Promoter in Goat

以内蒙古白绒山羊DNA为模板,参照牛的角蛋白14(keratin 14,K14)基因序列设计引物,PCR扩增获得长度为2834 bp(GenBank登录号:JQ063036)的片段,包括山羊K14基因启动子区2631 bp和结构基因第1外显子区203 bp。序列比对山羊K14基因启动子序列及其多态性分析结果发现,山羊K14基因启动子结构与人K14基因启动子不同,只含有Sox-5、GATA-1、GATA-1/2结合位点等3个共同的转录因子结合位点。但与牛K14基因启动子有较高的相似性,含有Sox-5、SRY、CdxA、MyoD、MZF1、GATA-1/3、Nkx-2结合位点等7个共同的转录因子结合位点;并发现山羊K14基因启动子区存在deltaE、GATA-2、GATA-1、Sp-1、AML-1a等5处独有的转录因子结合位点。此外,对山羊K14基因启动子进行多态性分析,发现4处单核苷酸多态位点,其中2处SNPs位于Nkx-2(-2004—-1996 bp)转录因子结合位点或Nkx-2(-1590—-1583 bp)临近几个碱基内。

According to the bovine keratin 14（K14） gene sequence designed the PCR primers,and acquired the 2834 bp gene sequence（GenBank accession：JQ063036） by the DNA of Inner Mongolia Cashmere goat.This region included 2631 bp promoter region and 203 bp the first exon of structural gene of goat K14 gene.Sequence aligenment and analysis,and study the polymorphism of K14 promoter.It was found that goat＇s and human＇s structure of K14 promoter were different,both only having three common transcription factor binding sites such as Sox-5,GATA-1,GATA-1/2 binding sites.However,with the cattle＇s had a higher similarity,containing the Sox-5,SRY,CdxA,MyoD,MZF1,GATA-1/3,Nkx-2 binding sites etc.7 common transcription factor binding sites.And it was found that goat K14 promoter region had five unique transcription factor binding sites,such as deltaE,GATA-2,GATA-1,Sp-1,AML-1a binding sites.In addition,the single nucleotide polymorphism（SNP） of goat K14 promoter was investigated,including four SNPs.Among these two novel SNPs were located in Nkx-2 binding site（-2004—-1996 bp） or near the several bases of Nkx-2 binding site Nkx-2（-1590—-1583 bp）.