猪精子黏合分子1（SPAM1）基因新SNPs的鉴别及其在12个中外猪中的遗传变异研究

精子黏合分子1 (SPAM1)是一种在受精过程中起重要作用的精子抗原,以10 个中外猪种的20个无遗传相关公猪个体的基因组DNA为模板,扩增猪SPAM1基因的部分第二外显子和第三内含子2 个DNA片段,经PCR产物直接测序并作序列比较分析后在外显子2和内含子3上分别鉴别到1个新的单核苷酸多态(SNPs)位点,其中外显子2上的T>A碱基颠换(M191T-A)可导致其编码的苯丙氨酸突变为酪氨酸.SMART软件分析显示M191T-A处在SPAM1蛋白糖基水解酶保守域的编码区.建立PCR- TasI -RFLP方法检测了4个西方商业猪种以及8 个中国地方猪种791个个体在M191T-A位点的遗传变异情况,结果在八眉猪、莱芜黑猪、米林藏猪、杭猪、剑河白香猪、大白猪、长白猪和杜洛克猪中检测到突变型等位基因A;而在二花脸猪、民猪、五指山猪和皮特兰猪中仅存在TT基因型,表现为极端的单态分布.

Identification of Two New SNPs in Porcine Sperm Adhesion Molecule 1(SPAM1)Gene and Its Genetic Variation Analysis in 12 Chinese and Western Pig Breeds

The sperm adhension molecule 1(SPAM1) as a sperm antigen,plays an important role in fertilization.Two novel single nucleotide polymorphisms(SNPs) were identified in exon 2 and intron 3 of the porcine SPAM1 gene by comparative sequencing using 20 unrelated boars DNA samples from 10 pig breeds.The T>A substitution at position 191 in the open reading frame(M191T-A),which led to the exchange of phenylalanine to tyrosine,was located in the conservative glycosyl hydrolyses domain by predicting with SMART software.The genetic variations at the M191T-A locus were investigated by PCR-TasI-RFLP on samples of 791 pigs from 4 western commercial pig breeds including Landrace,Large White,Duroc and Pietrain,and 8 Chinese indigenous pig breeds consisting of Bamei,Laiwu Black,Milin Tibetan,Hang,Jianhebai Xiang,Erhualian,Min and Wuzhishan pig.The results showed that the genetic polymorphisms at the SPAM1 M191T-A locus existed in Bamei,Laiwu Black,Milin Tibetan,Hang,Jianhebai Xiang,Landrace,Large White and Duroc breeds,whereas Erhualian,Min,Wuzhishan and Pietrain breeds only possessed the wild type TT.