月季eIF-5A基因的表达增强宿主大肠杆菌对高温和低温的耐性  

Increasing the Thermotolerance of E.coli with Expression of eIF-5A from Rosa chinensis

作　　者：石金磊[1] 徐健遥[2] 蒋昌华[2] 李敏[2] 李卓夫[1] 胡永红 明凤[2] 

机构地区：[1]东北农业大学农学院,哈尔滨150030 [2]复旦大学遗传所,上海200433 [3]上海市植物园,上海200231

出　　处：《中国生物工程杂志》2008年第1期18-24,共7页China Biotechnology

基　　金：国家自然科学基金(30570988);上海市科技兴农重点攻关项目[沪农科攻字(2005)第1-1-2号];新世纪优秀人才支持计划(NCET-07-0201),资助项目

摘　　要：真核生物翻译起始因子(eIF-5A)在调控生物生长发育、衰老与环境响应中起重要作用。设计eIF-5A基因的兼并引物,对月季受高温诱导的叶片cDNA进行PCR扩增,获得特异性片段,经回收、克隆和测序,确定该cDNA为月季eIF-5A(命名为RceIF5A),含有480bp的核苷酸,编码159个氨基酸。将该cDNA序列克隆到原核表达载体PET32a中,获得重组子pET32a-eIF5A。在高温(50℃)和低温(4℃)胁迫下含有该基因的大肠杆菌(Escherichiacoli)BL21(pET32a-eIF5A)比E.coliBL21(pET32a)抗性有明显的提高,据此认为含有重组子的E.coliBL21(pET32a-eIF5A)对高温和低温的抗性可能与eIF-5A基因的表达相关。该基因GenBank登录号为EF177192。Eukaryotie translation initiation factor 5A （ eIFSA ） is involved in cell proliferation, senescence, development and envirorimental response, eDNA of Chinese rose was amplified by PCR with degeneracy primer. The special fragment with 480 bp nucleic acid encoded a deduced amino acid sequence of 159 residues was obtained and named as RceIF-SA （GenBank No. EF177192）. The gene was cloned into pET32a-veetor and the recombination vector was named as pET32a -eIFSA. E. coli line BL21 （ pET32a -eIFSA ） had more obvious tolerance than E. coli BL21 （pET32a） under the high （50~C） and low （4℃）temperature. By all accounts, thermotolerance enhancing was attributed to the heterogenous expression of eIF-SA gene in host cell line.

关 键 词：EIF-5A 月季 原核表达 温度胁迫 

分 类 号：Q819[生物学—生物工程]