摘要
采用普通PCR和RACE技术克隆了西伯利亚鲟Acipenser baerii热休克蛋白HSP70 cDNA的全序列,该序列全长为2 343 bp,其中5'非编码区为140 bp,3'非编码区为256 bp,可读编码框(ORF)为1 947bp,编码为648个氨基酸。该氨基酸序列中含有HSP70家族的3个特征序列——IDLGTTYS、IFDLGGGTFD-VSIL和IVLVGGSTRIPKIQK,细胞质特征性保守序列为EEVD,C端重复序列为GGMP。该cDNA序列与其它生物的HSP70 cDNA序列一样具有很高的相似性。系统发育树显示,西伯利亚鲟与非洲爪蛙蟾Xenopus laevis、密西西比短吻鳄Alligator mississippiensis、美西螈Ambystoma mexicanum的亲缘关系较近。实时定量分析结果表明,水温为17.5℃时,西伯利亚鲟肝脏、鳃、脾脏、心脏、肌肉、中肠、性腺、脑8种组织中均有HSP70表达,其中HSP70在脾脏中的表达量最高,鳃中的次之,肝脏中最低(P<0.05)。
A full length cDNA encoding heat shock protein 70（HSP70） was cloned from liver RNA in Siberian sturgeon（Acipenser baerii）,and was found to contain 2 343 bp,including 5′untranslated region of 140 bp,3′untranslated region of 256 bp and an open reading frame of 1 947 bp.The deduced 648 amino acids sequence with estimated molecular of 71 000 and theoretical isoelectric point of 5.21 had high similarity with other vertebrate HSP70 genes in NCBI.Sequence analysis indicated that the HSP70 cDNA was consisted of three signature sequences IDLGTTYS,IFDLGGGTFDVSIL and IVLVGGSTRIPKIQK belonging to HSP70 family.Phylogenetic tree revealed that Alligator mississippiensis,Xenopus laevis,Ambystoma mexicanum and Sebrian Sturgeon showed a close genetic relationship.Using fluorescent real-time quantitative PCR,the HSP70 was expressed in the liver,gill,spleen,heart,intestine,muscle,gonad and brain of Siberian sturgeon,with the peak in the spleen and the minimum in the liver（P0.05）,indicating that the HSP70 cDNA could be used in understanding of anti-stress mechanism in Siberian sturgeon and in genetic breeding improvement.
出处
《大连海洋大学学报》
CAS
CSCD
北大核心
2012年第2期150-157,共8页
Journal of Dalian Ocean University
基金
农业部公益性行业科研专项(201003055-05)
北京市农林科学院青年基金资助项目(QNJJ201015)
