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采用RACE技术,对两个奥利亚罗非鱼群体(Oreochromis aurea)(美国奥利亚和中国奥利亚)热休克蛋白Hsp70基因完整编码区(code sequences,CDS)cDNA的进行克隆测序。序列分析结果表明:两个奥利亚罗非鱼群体热休克蛋白Hsp70基因CDS序列完全相同,全长1 923 bp,编码640个氨基酸,相对分子质量为70.29×103,理论等电点5.462,均具有Hsp70家族的3个签名序列:IDLGTTYS、IFDLGGGTFD、VVLVGGSTRIPKIQK;核定位信号标签KRKHKKDISQNKRALRR;Dank特征基序DLGTT-S-V;胞质Hsp70特征基序EEVD;靠近C端的GGMP4肽序列;2个糖基化位点NKSI和NVSA。对所得基因序列与已发表的青锵(Oryzias latipes)等物种Hsp70基因的氨基酸序列进行同源性比较,发现两个奥利亚罗非鱼群体与莫桑比克罗非鱼最高99.4%,与牙鲆最低83.9%;系统进化树分析表明奥利亚罗非鱼的Hsp70 cDNA序列与青鳉等物种的Hsp70基因聚在一支,而与牙鲆的Hsp70基因相分离。
The cDNA sequences of the heat shock protein Hsp70 gene coding sequences (CDS) from two populations of Oreochromis aurea (Oreochromis aurea and Oryza spp. In China) were cloned by RACE technique. The results of sequence analysis showed that the CDS sequences of Hsp70 gene from two populations of Tilia tritici were the same, with a total length of 1 923 bp, encoding 640 amino acids with a relative molecular mass of 70.29 × 103 and a theoretical isoelectric point of 5.462 Three signature sequences with Hsp70 family: IDLGTTYS, IFDLGGGTFD, VVLVGGSTRIPKIQK; nuclear localization signal tag KRKHKKDISQNKRALRR; Dank signature motif DLGTT-SV; cytoplasmic Hsp70 signature motif EEVD; GGMP4 peptide sequence near C-terminus; two glycosylation sites Point NKSI and NVSA. A comparison of homology between the obtained gene sequence and the published amino acid sequence of the Hsp70 gene of Oryzias latipes showed that the two O. aureus populations were the highest with the Mozambique tilapia at 99.4% Phylogenetic tree analysis showed that the Hsp70 cDNA sequence of O. tilapia was clustered with the Hsp70 gene of barley and other species and separated from the Hsp70 gene of Japanese flounder.