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.
出处
《四川大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第4期818-821,共4页
Journal of Sichuan University(Natural Science Edition)
作者简介
柴玉荣（1976-），女，2002级博士研究生，E-mail：yrchai@163．com
