CAJ | 학술논문

采用RT-PCR法从杂交鲟(♀Huso huso×♂Acipenser schrencki)和匙吻鲟(Polyodon spathula)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).
채용RT-PCR법종잡교심(♀Huso huso×♂Acipenser schrencki)화시문심(Polyodon spathula)간장RNA극륭획득HSP70기인적전장cDNA ( HSP70 cDNA).소측정적HSP70 cDNA서렬여NCBI/GenBank상등재적즉(GenBank No.DQ872648)동원성최고,잡교심화시문심분별체96%화98%,잡교심HSP70서렬위382 bp,시문심위334 bp.장잡교심화시문심여기타척추동물HSP70안기산서렬용DNAstar연건진행상사도비교,어류여포유동물、량서류비주조섬지간HSP70안기산서렬상사도평균치분별위86.2%화86.8%.어류지간적안기산서렬상사도교대,평균위93.8%,표현출교고적보수성.이HSP70핵감산서렬위분자표기,용MEGA4연건중최대간약법(MP)구건료12개물충HSP70계통발육수,식별출3개대적단계류군:잡교심、시문심、단두방、즉、리、반마어취위류군일(bootstrap 96);홍준화대서양해취위류군이(bootstrap 100);인류화갈가서류취위류군삼(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 (HSP70 cDNA) from hybrid sturgeon and Polyodon spathula were 382 bp and 334 bp long,respectively.Compared with other vertebrates' amino acid sequence of HSP70 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 major 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).