布莱凯特黑牛CoQ_(10)合成相关基因COQ2 cDNA克隆及其多态性分析  

Cloning of COQ2 gene cDNA and analyzing of its genetic diversity in the black cattle

作　　者：牛召珊[1,2] 董雅娟[1,2,3] 赵仕全[1,2] 杨莉[1,2] 张新浩[1,2] 柏学进[1,2,3] 

机构地区：[1]山东省黑牛繁育工程技术研究中心,山东青岛266109 [2]青岛农业大学动物胚胎工程中心,山东青岛266109 [3]山东布莱凯特黑牛科技股份有限公司,山东淄博256306

出　　处：《畜牧与兽医》2013年第10期31-35,共5页Animal Husbandry & Veterinary Medicine

基　　金：国家科技支撑计划(2012BAD39B05)

摘　　要：COQ2基因是真核生物中对羟基苯甲酸聚异戊二烯焦磷酸转移酶的编码基因,该酶是CoQ10生物合成过程的主要限速酶。试验根据已公布的普通牛COQ2基因序列和其cDNA序列,对COQ2基因的7个外显子区域进行引物设计。PCR产物克隆测序后,使用生物信息学软件对测序结果进行比对和拼接,得到布莱凯特黑牛COQ2基因cDNA序列,并对其外显子3进行了PCR-SSCP分析。结果表明:布莱凯特黑牛COQ2基因cDNA序列长1 546 bp,由7个外显子组成,氨基酸编码序列为第46 bp至1 161 bp处,编码371个氨基酸;与普通牛相比,布莱凯特黑牛PPT氨基酸第324位由天冬氨酸(N)突变为组氨酸(H)。SSCP检测发现外显子3中存在一个多态位点,在102 bp处发生A→T的碱基突变,产生了AA、AB、BB三种基因型,分析发现AB基因型频率最高,B等位基因频率(0.533)比C等位基因(0.467)高,为优势等位基因。该位点多态信息含量(PIC=0.374)为中度多态。该结果为进一步分析COQ2基因SNP位点与布莱凯特黑牛CoQ10合成情况及肉质性状的关联性奠定了基础,对布莱凯特黑牛品种培育具有重要的指导意义。COQ2 gene is coding the enzyme of p-hydroxybenzoic acid polyprenyl pyrophosphate transferase （PPT） in eukaryotes, which is the rate-limiting enzyme of the biosynthesis of Coenzyme Q10 （CoQ10） in vivo. According to the published bovine COQ2 gene sequence and cDNA sequence, the present study has designed seven pairs of primers for the seven exon regions. Using the bioinformatics software to compare and splice on the sequencing results, we did the cDNA sequence of Black Cattle COQ2 gene. PCR-SSCP was used to find out the diversity sites in exon 3 of COQ2 gene. The results showed that the length of COQ2 gene cDNA was 1546bp in Black Cattle was composed by 7 exons. The amino acid coding sequence was from the 46bp to the 1161bp, which could encode 371 amino acids. Compared with ordinary cattle, the 324th amino acid of Black Cattle was mutated from aspartic acid （N） to histidine （H）. SSCP analysis found a polymorphism in the exon 3 of COQ2 gene in black Cattle, which made a mutation A to T in the 102th base. There were three genotypes AA, AB and BB, the frequency of AB genotype was the highest in the population. The frequency of B allele （0. 533） was higher than the frequency of A allele （0. 467） and was the dominant allele. The PIC was 0. 374 and belonged to moderate polymorphism. Our result provided a foundation for the analysis of the relationship between the COQ2 polymorphisms and CoQ10 biosynthesis and even the relationship with quality traits.

关 键 词：布莱凯特黑牛 CoQ10 COQ2基因 CDNA PCR—SSCP 

分 类 号：S813.23[农业科学—畜牧学]