| 程保平1、鹿连明2、彭埃天1、赵弘巍3、宋晓兵1、陈霞1.柑桔黄龙病菌外膜蛋白的原核表达与纯化[J].广东农业科学,2014,41(10):132-134 |
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柑桔黄龙病菌外膜蛋白的原核表达与纯化 |
| Prokaryotic expression and purification of the CandidatusLiberibacter asiaticus Omp protein |
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| DOI： |
| 中文关键词: 柑桔黄龙病菌 omp 基因 原核表达 蛋白纯化 |
| 英文关键词: Candidatus Liberibacter asiaticus omp gene prokaryotic expression protein purification |
| 基金项目:广东省农业科技重大技术研究项目（201201127）；
广东省现代农业产业技术体系建设专项（粤农[2009]380 号）；广
东省农业科学院院长基金（201305）；广东省科技创新人才出国
培养计划（粤农科[2013]74 号） |
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| 中文摘要: |
| 为获得大量高纯度的柑桔黄龙病菌Omp 外膜重组蛋白、应用PCR 方法从染病柑桔模板中扩增出黄龙
病菌omp 基因的3 个片段、将其克隆到pMDl9-T 载体、再亚克隆到pET32a 原核表达载体、构建pET32a-omp 原核表
达重组质粒、然后转化大肠杆菌BL21（DE3）、经IPTG 诱导并用SDS-PAGE 电泳检测带组氨酸标签的Omp 融合蛋白
表达后、用镍柱分离纯化该融合蛋白。结果表明院柑桔黄龙病菌omp 基因序列的两个片段在37益经1 mmol/L IPTG
诱导后、在大肠杆菌得到了高效表达；镍柱纯化蛋白得到预期大小（约55 ku）的融合蛋白。 |
| 英文摘要: |
| In order to get highly purified Omp protein of Candidatus Liberibacter asiaticus for function research, three
fragments of the omp gene were amplified from infected citrus template with PCR. Then the three fragments were cloned
into pET32a vector to construct pET32a -omp prokaryotic expression recombinant plasmid. The plasmid were then
transformed into E. coli cell BL21(DE3). After that, Omp (His tag) fusion protein was induced with IPTG and detected with
SDS-PAGE electrophoresis. In the end, Omp fusion protein was purified with nickel column. The result showed that two
fragments of the Omp protein were efficiently expressed in E. coli, after induced with IPTG on 37益. Expected protein
(about 55 ku) were gotten after purification with nickel column. |
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