摘要
生物素羧基载体蛋白(biotin carboxyl carrier protein,BCCP)负责将羧基从生物素羧化酶转移到羧基转移酶上,对于长链脂肪酸的从头合成与种子含油量的形成具有十分重要的作用。以大肠杆菌E.coli DH5α的DNA为模板,根据大肠杆菌基因组的Genbank中的核苷酸序列,设计2条特异引物进行扩增,克隆了大肠杆菌乙酰辅酶A羧化酶(Acetyl CoA Carboxylase,ACCase)的生物素羧基载体蛋白亚基基因accB,构建accB的原核表达载体pGEX-4T-accB,转化大肠杆菌BL21(DE3)工程菌株,IPTG诱导进行融合表达,成功诱导表达了GST-accB融合蛋白,大小为43 kDa,该基因的原核表达为蛋白的纯化和进一步研究其功能及应用打下了良好的基础。
Acetal-CoA Carboxylase （ACCase） catalyzes the first step in de novo biosynthesis of fatty acids. Bio- tin carboxyl carrier protein （BCCP）, as one of the subunits of plastidial ACCase, controls the transfer of carbox- yl from acetyl-CoA carboxylase to carboxyhransferase. Special primer designed and synthesized based on the sequence of the codon of E. coli, and the gene of accB was obtained by PCR amplification and cloned into pro- karyotic expression vector pGEX-4T-1. Then the recombinant expression plasmid pGEX-4T-accB was trans- formed into E. coli BL21 and the fusion protein of accB was produced by IPTG induction. The analysis of SDS-PAGE showed that the fusion pGEX-4T-accB protein was expressed about 43 kD.
出处
《中国农学通报》
CSCD
北大核心
2009年第23期74-77,共4页
Chinese Agricultural Science Bulletin
基金
浙江省自然科学基金项目"异质型乙酰辅酶A羧化酶(ACCase)各亚基分别和组合定域油菜籽粒质体的高油作用研究"(No.Y305251)
