集胞藻PCC6803基因sll0853的克隆、优化表达及其结构功能预测  

Cloning,Optimization of Prokaryotic Expression of sll0853 and Prediction of the Structure and Function

作　　者：刘欣[1] 陈思礼[1] 梅菊[1] 尤隽丹[1] 

机构地区：[1]中南民族大学生命科学学院,湖北武汉430074

出　　处：《山西大学学报（自然科学版）》2012年第3期552-556,共5页Journal of Shanxi University(Natural Science Edition)

基　　金：国家自然科学基金(31001099/C190101);湖北省全球基金艾滋病项目(SZY10008);微生物与生物转化中南民族大学重点实验室项目(XJS09002)

摘　　要：从集胞藻(Synechocystis sp.)PCC6803提取总DNA,利用PCR扩增目的基因sll0853,构建重组T-0853克隆载体和pET-0853原核表达载体.为了提高sll0853在大肠杆菌E.coli BL21(DE3)中的表达量,通过改变诱导温度、诱导时间及诱导剂浓度等条件对表达量产生影响,以SDS-PAGE电泳分析证明sll0853基因蛋白表达的最佳条件.结果表明:目的蛋白在28℃、0.2mmol/L IPTG、诱导6h表达量分别达高峰.通过生物信息学软件预测基因sll0853可能具有裂合酶的功能.The total DNA was extracted from Synechocystis sp.PCC6803,and sll0853 gene was screened and amplified by PCR.The sll0853 gene was connected with cloning vector,pMD18-T and a prokaryotic expression vector,pET-30a.In order to improve the expression level of sll0853 in E.coli BL21(DE3),the optimal conditions for expression of fusion protein were analyzed by SDS-PAGE at different concentration,temperature and time of IPTG.The results showed that the fusion protein expression of recombinant plasmid in E.coli BL21(DE3) was the best at 0.2 mmol/L of IPTG for 6 h at 28 ℃.In addition,the protein of sll0853 maybe have nature of CpeS through the prediction of bioinformatics.

关 键 词：集胞藻PCC6803 sll0853 克隆表达 结构功能预测 

分 类 号：Q943[生物学—植物学]