杂交鲟和匙吻鲟HSP70 cDNA克隆与序列分析  

Cloning and Sequence Analysis of HSP70 cDNA from Hybrid Sturgeon and Paddle Fish(Polyodon spathula)

作　　者：高宇[1] 袁改玲[1] 李大鹏[1] 陈昌福[1] 

机构地区：[1]华中农业大学水产学院,武汉430070

出　　处：《华中农业大学学报》2010年第1期85-89,共5页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金项目(30970529);家畜疫病病原生物学国家重点实验室(中国农业科学院兰州兽医研究所)开放课题资助

摘　　要：采用RT-PCR法从杂交鲟(♀Huso huso×♂Acipenserschrencki)和匙吻鲟(Polyodonspathula)肝脏RNA克隆获得HSP70基因的全长cDNA(HSP70 cDNA)。所测定的HSP70 cDNA序列与NCBI/GenBank上登载的鲫(GenBank No.DQ872648)同源性最高,杂交鲟和匙吻鲟分别达96%和98%,杂交鲟HSP70序列为382 bp,匙吻鲟为334 bp。将杂交鲟和匙吻鲟与其它脊椎动物HSP70氨基酸序列用DNAstar软件进行相似度比较,鱼类与哺乳动物、两栖类非洲爪蟾之间HSP70氨基酸序列相似度平均值分别为86.2%和86.8%。鱼类之间的氨基酸序列相似度较大,平均为93.8%,表现出较高的保守性。以HSP70核苷酸序列为分子标记,用MEGA4软件中最大简约法(MP)构建了12个物种HSP70系统发育树,识别出3个大的单系类群:杂交鲟、匙吻鲟、团头鲂、鲫、鲤、斑马鱼聚为类群一(bootstrap 96);虹鳟和大西洋鲑聚为类群二(bootstrap 100);人类和褐家鼠类聚为类群三(bootstrap 89)。The heat shock protein 70 （HSP70） complete cDNAs were cloned from hybrid sturgeon （ ♀ Huso huso × ♂ Acipenser schrencki ） and Polyodon spathula. The results of the hybrid sturgeon and Polyodon spathula ＇s sequence showed that this nucleotide sequence of HSP70 gene has 96% and 98% identity with the Carassius auratus （GenBank No. DQ872648. 1） sequence published by NCBI/GenBank. The full length cDNA （HSPT0 cDNA） from hybrid sturgeon and Polyodon spathula were 382 bp and 334 bp long,respectively. Compared with other vertebrates＇ amino acid sequence of HSPT0 by DNAstar software,the sequence similarity was 86.2% between fish and mammals, and 86. 8% between fish and amphibian, respectively. The sequence similarity was high among different fish species, and the average value was 93.8%,which shows a higher conservatism. Phylogenetic tree was constructed for 12 species based on HSP70 amino acid sequences using MP （maximum parsimony） method. Three maior clusters were recognized：cluster one consisted of hybrid sturgeon, Polyodon spathula ,Megalobrama amblycephala ,Carassius auratus,Cyprinus carpio and Danio rerio （bootstrap 96） ; cluster two consisted of Oncorhynchus mykiss and Salmo salar （bootstrap 100）; and cluster three consisted of Homo sapiens and Rattus norvegicus （bootstrap 89）.

关 键 词：热应激蛋白HSP70 杂交鲟 匙吻鲟 克隆 序列分析 

分 类 号：Q959.463[生物学—动物学]