5个绵羊品种BMP6基因部分片段的多态及序列分析  

Polymorphism and Sequence Analysis of Partial Fragment of BMP6 Gene in Five Sheep Breeds

作　　者：肖朝庭[1,2] 狄冉[1] 储明星[1] 傅衍[2] 方丽[1] 马月辉[1] 李奎[1] 徐作鹏 

机构地区：[1]中国农业科学院北京畜牧兽医研究所农业部畜禽遗传资源与利用重点开放实验室,北京100193 [2]浙江大学动物科学学院,杭州310029 [3]河北省平泉县农牧局,平泉067500

出　　处：《畜牧兽医学报》2008年第12期1631-1639,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家科技基础条件平台建设计划(2005DKA21101);国家973计划(2006CB102105);国家自然科学基金项目(30300248);北京市自然科学基金项目(5042017)资助

摘　　要：根据GenBank发表的绵羊骨形态发生蛋白6(Bone morphogenetic protein 6,BMP6)基因部分序列所包含的外显子5、6、7和小鼠BMP6基因外显子5、6、7序列设计3对引物,采用PCR-SSCP技术检测BMP6基因外显子5、6和7在小尾寒羊、湖羊、多赛特、特克塞尔、考力代5个绵羊品种301个个体中的单核苷酸多态性。结果发现这3对引物的扩增片段在检测的5个品种中均无多态性,说明所检测的BMP6外显子5、6、7序列比较保守。同时克隆了小尾寒羊(695 bp)和济宁青山羊(699 bp)部分BMP6 mRNA以及小尾寒羊BMP6基因外显子3-4的内含子序列和两端部分外显子序列(785 bp)。发现小尾寒羊和济宁青山羊的核苷酸和氨基酸序列差异很小,两者的核苷酸序列同源性高达99.28%,氨基酸序列同源性为98.1%,除济宁青山羊中插入一个丙氨酸外,两者只有4个氨基酸的差异。绵羊与牛、人、小鼠和大鼠的核苷酸同源性分别为97.04%、81.82%、84.68%和85.06%;氨基酸序列同源性分别为99.05%、91.43%、90.48%和92.38%,均大于90%,表明各物种BMP6核苷酸序列虽然差异较大,但氨基酸序列却非常保守。Single nucleotide polymorphisms （SNPs） of exons 5, 6, 7 of bone morphogenetic protein 6 （BMP6） gene were detected in 301 individuals of five sheep breeds （Small-tailed Han, Hu, Dorset, Texel and Corriedale sheep） with three pairs of primers by polymerase chain reaction （PCR）-single strand conformation polymorphism （SSCP）. No polymorphisms were detected in the amplified regions of three pairs of primers in five sheep breeds tested, which may indicate that the sequences of exons 5, 6, 7 of BMP6 gene were rather conserved. On the other hand, partial sequences of BMP6 mRNA were cloned from Small-tailed Han sheep （695 bp） and Jining Grey goat （699 bp） respectively, and the sequences （785 bp） of intron between exon 3 and exon 4 with some of the two flanking exons were also sequenced in Small-tailed Han sheep. The two BMP6 sequences show high identity with 99.28% at nucleotide level and 98.1% at amino acid level, and besides an insert of alanine in goat, only 4 amino acid alterations were found between them （211 in total）. Compared with the ovine nucleotide and amino acid sequence of BMP6 with that of cattle, human, mouse and rat, identities of 97.04%, 81.82%, 84. 68%, 85.06% and identities of 99.05%, 91.43%, 90.48%, 92.38% were revealed respectively. The results suggested that although the nucleotide changes are a little big, the amino acid sequences of BMP6 are very conservative among species.

关 键 词：绵羊 骨形态发生蛋白6基因 PCR-SSCP 

分 类 号：S826.2[农业科学—畜牧学]