水稻中一个编码18.0 kDa I类低分子量热激蛋白的cDNA的克隆与特性分析

Cloning and characterization of a cDNA encoding an 18.0-kDa class-I low-molecular-weight heat-shock protein from rice.

作者信息

Lee Y L, Chang P F, Yeh K W, Jinn T L, Kung C C, Lin W C, Chen Y M, Lin C Y

机构信息

Department of Botany, National Taiwan University, Taipei, Taiwan.

出版信息

Gene. 1995 Nov 20;165(2):223-7. doi: 10.1016/0378-1119(95)00562-k.

DOI:10.1016/0378-1119(95)00562-k

PMID:8522180

Abstract

A novel cDNA clone, Oshp18.0 cDNA, encoding a rice (Oryza sativa L. cv. Tainong 67) 18.0-kDa heat-shock protein (HSP), was isolated from a cDNA library of heat-shocked rice seedlings by use of the rice HSP cDNA, Oshsp17.3 cDNA, as a probe. The sequence showed that Oshsp18.0 cDNA contains a 749-bp insert encoding an ORF of 160 amino acids, with a predicted molecular mass of 18.0 kDa and a pI of 7.3. Sequence comparison reveals that Oshsp18.0 cDNA is highly homologous to other low-molecular-weight (LMW) HSP cDNAs. Also, the results of hybrid-selected in vitro translation clearly establish that Oshsp18.0 cDNA is the rice 18.0-kDa LMW HSP-encoding cDNA clone. The recombinant Oshsp18.0 fusion protein produced in Escherichia coli was of the size predicted, and was recognized by the class-I rice 16.9-kDa HSP antiserum. The results suggest that Oshsp18.0 cDNA is an 18.0-kDa class-I LMW HSP- encoding cDNA clone from rice.

摘要

利用水稻热激蛋白cDNA（Oshsp17.3 cDNA）作为探针，从热激处理的水稻幼苗cDNA文库中分离出一个新的cDNA克隆，即Oshp18.0 cDNA，它编码一种水稻（Oryza sativa L. cv. Tainong 67）18.0 kDa的热激蛋白（HSP）。序列分析表明，Oshsp18.0 cDNA包含一个749 bp的插入片段，编码一个由160个氨基酸组成的开放阅读框，预测分子量为18.0 kDa，等电点为7.3。序列比较显示，Oshsp18.0 cDNA与其他低分子量（LMW）HSP cDNA高度同源。此外，体外杂交选择翻译的结果清楚地表明，Oshsp18.0 cDNA是水稻18.0 kDa LMW HSP编码的cDNA克隆。在大肠杆菌中产生的重组Oshsp18.0融合蛋白大小与预测一致，并被I类水稻16.9 kDa HSP抗血清识别。结果表明，Oshsp18.0 cDNA是来自水稻的一个编码18.0 kDa I类LMW HSP的cDNA克隆。