亚洲黑熊四川亚种(Ursus thibetanus mupinensis)线粒体ATP合酶和亚基基因克隆及序列分析  

Cloning and Sequence Analysis of mtDNA ATP Synthase F_0 Subunit 8(ATP8) and Subunit 6(ATP6) Gene from Asian Black Bear Sichuan Subspecies(Ursus thibetanus mupinensis)

作　　者：郝彦哲[1] 杜玉杰[1] 吴夏[1] 张田[1] 侯万儒[1] 

机构地区：[1]西华师范大学生命科学学院,四川南充637002

出　　处：《重庆师范大学学报（自然科学版）》2008年第1期20-24,共5页Journal of Chongqing Normal University:Natural Science

基　　金：国家自然科学基金项目(No.30470261);四川省教育厅重点科研项目(No.402044);西华师范大学科研启动项目(2003);四川省重点学科建设项目(No.SZD0420)

摘　　要：根据已报道的部分哺乳动物线粒体ATP合酶F0亚基的相关信息设计引物,运用PCR技术,首次从亚洲黑熊四川亚种(Ursus thibetanus mupinensis)的肌肉组织总DNA中成功克隆了线粒体ATP合酶F0亚基8(ATP8)和亚基6(ATP6)的序列,并对其进行了初步分析。结果表明:PCR扩增产物的总长度为942 bp,其中842 bp为四川黑熊ATP8和ATP6基因的编码区。ATP8和ATP6基因存在一段长43 bp的重叠区域。ATP8基因长204 bp,编码67个氨基酸残基的蛋白质,其蛋白分子质量为7.9 KD,等电点为10.35;ATP6基因长682 bp,编码226个氨基酸残基的蛋白质,其蛋白分子质量为24.8 KD,等电点为10.63。四川黑熊线粒体ATP合酶F0亚基8和亚基6与其他已报道的部分哺乳动物具有很高的同源性。以基因序列为数据构建的进化树表明四川黑熊和美洲黑熊的亲缘关系最近。本研究为在分子水平上探究黑熊线粒体基因组的遗传特点,探究物种进化关系和物种多样性提供了科学参考。Mitochondrial ATP synthase F0 subunit 8（ATP8） and subunit 6（ATP6） gene in muscular tissue of Asian black bear Sichuan subspecies（Ursus thibetanus mupinensis） are amplified for the first time through PCR technology with primers designed according to ATP8 genes and ATP6 genes of some other mammals reported ,which are cloned,sequenced and analyzed preliminarily.The result shows that the full length of the PCR product is 942bp which contains ORF 842 encoding the ATP8 and ATP6 genes with a 43bp overlap between them.The ATP8 sequence is 204bp encoding a protein with 67 amino acid residues whose molecular weight is 7.9KD and isoelectric point is 10. 35. The ATP6 gene is 682bp encoding a protein with 226 amino acid residues whose molecular weight is 24. 8KD and isoelectric point isl0.63. Both of them are highly homologous to those of other mammals reported. Construction of phylogenetic tree based on nucleotide sequence manifests that the Asian black bears with Sichuan subspecies have closer relationship to the American black bears.

关 键 词：亚洲黑熊亚种 线粒体 ATP合酶 亚基 基因克隆 四川 

分 类 号：Q959.838[生物学—动物学] Q343.17