Gade Nitin, Mahapatra R K, Sonawane Arvind, Singh V K, Doreswamy Ramesh, Saini Mohini
Division of Physiology and Climatology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, 243 122, India.
Mol Biol Int. 2010;2010:108429. doi: 10.4061/2010/108429. Epub 2010 Jun 29.
Heat shock protein 70 (HSP 70) plays a vital role by bestowing cytoprotection against diverse kinds of stresses. The ubiquitous HSP 70 proteins are the most abundant and temperature sensitive among all the HSPs. The present paper has characterized HSP70-1 cDNA in goat (Capra hircus). Total RNA isolated from goat peripheral blood mononuclear cells was reverse transcribed to cDNA that was used for amplification of HSP 70-1 gene. PCR product (1926 bp) was cloned in pGEM-T easy vector and sequenced. Sequence analysis revealed 1926-bp-long open reading frame of HSP 70-1 gene encoding 641 amino acids in goat, as reported in cattle. At nucleotide level, goat HSP 70-1 was found to be 96-99% similar to that of sheep (partial), cattle, and buffalo whereas the similarity at amino acid level was 95-100%. Nonsynonymous substitutions exceeding synonymous substitutions indicate the evolution of this protein through positive selection among domestic animals. Goat and sheep appear to have diverged from a common ancestor in phylogenetic analysis. Predicted protein structures of goat HSP 70 protein obtained from deduced amino acid sequence indicated that the functional amino acids involved in chaperoning through ATPase hydrolytic cycle and in uncoating of clathrin coated vesicles are highly conserved.
热休克蛋白70（HSP 70）通过赋予细胞对多种应激的保护作用发挥着至关重要的作用。普遍存在的HSP 70蛋白在所有热休克蛋白中最为丰富且对温度敏感。本文对山羊（Capra hircus）中的HSP70-1 cDNA进行了表征。从山羊外周血单核细胞中分离的总RNA被逆转录为cDNA，用于扩增HSP 70-1基因。PCR产物（1926 bp）被克隆到pGEM-T easy载体中并进行测序。序列分析显示，如在牛中报道的那样，山羊HSP 70-1基因的开放阅读框长1926 bp，编码641个氨基酸。在核苷酸水平上，发现山羊HSP 70-1与绵羊（部分）、牛和水牛的相似性为96%-99%，而在氨基酸水平上的相似性为95%-100%。非同义替换超过同义替换表明该蛋白在家畜中通过正选择进化。在系统发育分析中，山羊和绵羊似乎从一个共同祖先分化而来。从推导的氨基酸序列获得的山羊HSP 70蛋白的预测蛋白质结构表明，通过ATPase水解循环参与伴侣作用以及在网格蛋白包被小泡去包被过程中起作用的功能性氨基酸高度保守。
