光棘球海胆CYP17基因的SNPs标记筛选及遗传分析  

SNPs screening and genetic analysis of CYP17 gene in sea urchin Strongylocentrotus nudus

作　　者：朱琳琳[1,2] 王秀利[1,2] 王洪迪[2] 郭晓黎[2] 常亚青[1,2] 刘洋[1] 仇雪梅[1,2] 

机构地区：[1]大连海洋大学农业部北方海水增养殖重点实验室,辽宁大连116023 [2]大连海洋大学水产与生命学院,辽宁大连116023

出　　处：《大连海洋大学学报》2013年第4期373-377,共5页Journal of Dalian Ocean University

基　　金：国家“863”计划重大项目(2012AA10A412);国家自然科学基金资助项目(31101923);辽宁省自然科学基金资助项目(20102043);辽宁省百千万人才工程项目(2010921093);大连海洋大学科研项目(2012HYDX02)

摘　　要：采用聚合酶链式反应、单链构象多态性(PCR-SSCP)技术和基因测序方法筛选光棘球海胆Strongylocentrotus nudus细胞色素17α羟化酶/17,20裂解酶(17α-hydroxylase/17,20-lyase,CYP17)基因编码区单核苷酸多态性(SNPs),并进行了基于SNPs的遗传分析。结果表明:在CYP17基因的270、417、666 nt处均发生了碱基的点突变,即G270A、A417G和C666T,前两个突变位点位于引物对BF/BR的扩增产物中,后一个突变位点位于引物对CF/CR的扩增产物中;引物对BF/BR的扩增产物可形成3种基因型,即AA、BB和AB基因型,A和B等位基因频率分别为0.54和0.46,AA、AB和BB基因型频率分别为0.30、0.48和0.22,该座位的杂合度(He)、纯合度(Ho)、多态性信息含量(PIC)和有效等位基因数(Na)分别为0.496 8、0.503 2、0.373 4和1.987 3;引物对CF/CR的扩增产物形成了两种基因型,即CC和CD基因型,C和D等位基因频率分别为0.68和0.32,CC和CD基因型频率分别为0.84和0.16,该座位的He、Ho、PIC和Na分别为0.268 8、0.731 2、0.232 7和1.367 6。遗传变异分析结果表明:两个座位均属于低度多态性,而且群体遗传杂合度较低,这反映了该群体的遗传一致性较高。本研究筛选的3个SNPs为今后CYP17基因在光棘球海胆性腺生长发育研究上提供了前期科研基础。SNPs of 17ot-hydroxylase/17,20-1yase gene were detected by PCR-single strand conformation polymor- phism and genetic diversity was analyzed based on the SNPs in sea urchin Strongylocentrotus nudus. The results showed that there were three SNPs in the 270 nt nucleotide, the 417 nt nucleotide and the 666 nt nueleotide of open reading frame of CYP17 eDNA, namely G270A,A417G and C666T. The first tow mutations were found in primer pairs BF/BR, and the last mutation was located in primer pairs CF/CR. The PCR production of primer pairs BF/ BR formed genotypic AA, BB and AB, with frequency of 0.54 in Allele A and 0.46 in Allele B. The frequencies were found 0.30 in genotypic AA, 0.48 in genotypic BB and 0.22 in genotypic AB. The primer pairs BF/BR re- vealed that the heterozygosity was 0. 496 8, homozygosity 0.503 2, polymorphism information content 0. 373 4 and 1. 987 3 allele number of the locus. Similarly, the PCR production of primer pairs CF/CR formed genotypic CC and CD with the frequencies of 0.68 in Allele C and 0.32 in Allele D. The genotypic CC had frequency of 0.84 and the CD showed requency of 0.16. By the primer pairs CF/CR, there were the heterozygosity of 0.268 8, homozy- gosity of 0.731 2, polymorphism information content of 0. 232 7 and the locus allele number of 1. 367 6. The ge- netic analysis indicated that the two loci belonged to lower PIC, and the population heterozygosity was lower, with higher genetic consistency. The three SNPs are the base for further study of CYP17 gene in the sea urchin growth and development in future.

关 键 词：光棘球海胆 CYP17基因 单核苷酸多态性 遗传分析 

分 类 号：S917[农业科学—水产科学]