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| 幽门螺杆菌肽脱甲酰基酶基因的表达、纯化与活性检测 | |
| 引用本文： | 幺山山,曾明,章金刚.幽门螺杆菌肽脱甲酰基酶基因的表达、纯化与活性检测[J].药品评价,2005,2(3):184-186,190. |
| 作者姓名： | 幺山山 曾明 章金刚 |
| 作者单位： | 1. 中国药品生物制品检定所,北京,100050;军事医学科学院野战输血研究所,北京,100850 2. 中国药品生物制品检定所,北京,100050 3. 军事医学科学院野战输血研究所,北京,100850 |
| 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。 |
| 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 |
| 文章编号： | 1672-2809（2005）03-0184-04 |
| 收稿时间： | 2005-05-09 |
| 修稿时间： | 2005-06-15 |
Expression, purification of Helicobacter pylori peptide deformylase in E.coli and its activity determination |
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| YAO Shan-shan,ZENG Ming,ZHANG Jin-gang.Expression, purification of Helicobacter pylori peptide deformylase in E.coli and its activity determination[J].Drug Evaluation,2005,2(3):184-186,190. | |
| Authors: | YAO Shan-shan ZENG Ming ZHANG Jin-gang |
| Abstract: | Objective To develop new drugs targeting against peptide deformylase (PDF) of Helicobacter pylori, it is important to prepare a soluble and active PDF in E.coli. Method In this work, PDF gene (def) was cloned into pGEM-T easy vector and transferred to expression vector pET-32a. E. coli 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. |
| Keywords: | Helicobacter pylori Peptide deformylase gene expression gene amplification actiuity determination |
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