幽门螺杆菌肽脱甲酰基酶基因的表达、纯化与活性检测  

Expression, purification of Helicobacter pylori peptide deformylase in E.coli and its activity determination

作　　者：幺山山[1] 曾明[1] 章金刚[2] 

机构地区：[1]中国药品生物制品检定所,北京100050 [2]军事医学科学院野战输血研究所,北京100850

出　　处：《药品评价》2005年第3期184-186,190,共4页Drug Evaluation

摘　　要：目的高效表达幽门螺杆菌肽脱甲酰基酶(PDF)蛋白。方法以PCR方法克隆肽脱甲酰基酶基因(def),构建了融合表达载体pET-32a-def,在宿主菌BL21中进行了诱导表达。结果重组产物获高效表达,部分产物以可溶状态存在,经纯化后具有肽脱甲酰基酶活性。结论为PDF特性分析及PDF抑制剂筛选奠定了基础。Objective To develop new drugs targeting against peptide detormylase cobacter pylori, it is important to prepare a soluble and active PDF in E.coli. （PDF） ol Hell- Method In this work, PDF gene （det） was cloned into pGEM-T easy vector and transferred to expression vector pET-32a. E. coil cells BL21（DE3） carrying recombinant plasmid of pET-32a-def were induced by IPTG to express PDF. Result BL21 （DE3） carrying this plasmid produced high level of fusion protein, with molecular weight of 36kDa determined by SDS-PAGE. Most of the recombinant proteins were expressed as inclusion body, and only small part was soluble. Purified soluble recombinant PDF（rpET-PDF） shew peptide deformylase activity in vitro. Conclusions This work provided the basis for screening of PDF inhibitor.

关 键 词：幽门螺杆菌 肽脱甲酰基酶 基因表达 基因扩增 活性检测 

分 类 号：Q78[生物学—分子生物学]