盐藻rbcS基因克隆及其原核表达(英文)  

Cloning and Prokaryotic Expression of rbcS Gene from Dunaliella salina

作　　者：柴玉荣[1] 王亚锋[1] 王天云[1] 侯卫红[1] 王宁[1] 王建民[1] 薛乐勋[1] 

机构地区：[1]郑州大学医学院细胞生物研究室,郑州450052

出　　处：《四川大学学报（自然科学版）》2005年第4期818-821,共4页Journal of Sichuan University(Natural Science Edition)

摘　　要：1,5-二磷酸核酮糖羧化酶/加氧酶(Ribulose1,5-bisphosphatecarboxylase/oxygenase,E.C.4.1.1.39简称RuBisCO)是光合作用中的关键酶,作者应用RT-PCR技术从一种极其耐盐的生物—杜氏盐藻的总RNA中克隆RuBisCO的小亚基(rbcS)的cDNA,它的开放读码框为570bp,编码190个氨基酸.将此cDNA定向克隆于原核表达载体pET-30a(+)中,构建重组质粒pET-rbcS,经IPTG诱导,在大肠杆菌株BL21中表达.SDS-PAGE电泳表明,rbcS融合蛋白分子量约为26kDa.A full-length eDNA encoding the small subunit （rbcS） of ribulose-1,5-bisphosphate carboxylase/oxygenase （RuBisCO）from Dunaliella salina, one of the most halotolerant unicellular eukaryotic alga, was cloned by the method of reverse transcription polymerase chain reaction （RT-PCR）. The open reading frame of cloned fragment of cDNA for rbcS was 570 bp in length encoding 190 amino acids. The prokaryotic expression vector pET-rbcS was constructed by inserting the eDNA for rbcS into the pET-30a（＋） and transformed into E. coli BL21. The expressed fusion protein was analyzed by SDS-PAGE and a new specific band with molecular weight of about 26 kDa was found when induced by IPTG.

关 键 词：盐藻 1 5-二磷酸核酮糖羧化酶/加氧酶 RBCS 原核表达 

分 类 号：Q78[生物学—分子生物学]