| 罗晓庆,孙济宇,王慧,王晓波,王琪,蒋瑞雪.铜绿假单胞菌外膜蛋白OprH原核表达载体的构建与表达[J].中国烧伤创疡杂志,2014,(3):195~198. |
| DOI： |
| 中文关键词: 铜绿假单胞菌 外膜蛋白 原核表达载体 重组质粒 |
| 英文关键词:Pseudomonas aeruginosa Outer membrane protein Prokaryotic expression vector Recombinant plasmid |
| 基金项目:齐齐哈尔市科技局指令性科研计划课题（项目编号：SF-201106） |
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| 中文摘要: |
| 目的 克隆铜绿假单胞菌外膜蛋白OprH基因, 构建其原核表达载体并鉴定其表达? 方法 从铜绿假单胞菌中提取基因组DNA进行PCR扩增OprH基因; 采用T-A克隆技术构建pMD19T-OprH重组质粒, 并经酶切和序列测定后, 获得OprH片段插入到原核表达载体pET28b中, 以构建pET28b-OprH重组表达质粒, 并在表达宿主菌E.Coli BL21中经IPTG诱导表达及通过SDS-PAGE电泳鉴定? 结果 pET28b-OprH原核表达载体构建成功; 重组表达质粒经IPTG诱导后表达外膜蛋白OprH? 结论 成功克隆了OprH基因并获得原核表达物, 为进一步研究快速检测该菌的方法奠定了基础? |
| 英文摘要: |
| Objective To clone the gene of outer membrane protein OprH of Pseudomonas aeruginosa( PA) , construct a prokaryotic expression vector and identify its expression．Methods The DNA genome was extracted from PA, the OprH gene was amplified by primers of PCR. The recombinant plasmid pMD19T - OprH was constructed by using T - A cloning. After enzyme digestion and sequence analysis, the OprH gene was inserted into prokaryotic expression vector pET28b to construct recombinant expression plasmid pET28b -oprH, which was expressed in E.coli BL21 (DE3) cells with induction by IPTG. Then the expressed protein was analyzed by SDS-PAGE. Results Construction of the prokaryotic expression vector pET28b- oprH was successful. Then the recombinant expression plasmid expressed a corresponding protein OprH after being induced by IPTG. Conclusion OprH gene was successfully cloned and the prokaryotic expression product was obtained. It provides the basis for investigating a method of rapid detection of PA. |
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