›› 2010, Vol. 37 ›› Issue (12): 55-59.

• 生物技术 • 上一篇    下一篇

乳酸菌素基因PlnEPlnF在大肠杆菌中的异源表达

杨芳,余占桥,张日俊   

  1. (中国农业大学饲料生物技术实验室,动物营养学国家重点实验室,北京 100193)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-12-20 发布日期:2010-12-20
  • 通讯作者: 张日俊

Heterologly Expression of Plantaricin GenesPlnE and PlnF in E.coli

YANG Fang,YU Zhan-qiao,ZHANG Ri-jun   

  1. (Laboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, China Agricultural University, Beijing 100193, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-12-20 Published:2010-12-20
  • Contact: ZHANG Ri-jun

摘要: 为获得高产量纯品植物乳杆菌素PlnE和PlnF,研究其开发生物兽药的可能,试验通过人工合成植物乳杆菌素基因PlnEPlnF,构建表达载体pGEX-4T-E和pGEX-4T-F,经IPTG诱导表达条件优化后,进行亲和层析纯化融合表达蛋白,并使用SDS-PAGE进行鉴定。试验结果表明,pGEX-4T-E和pGEX-4T-F构建成功,纯化后获得2种产量高达30%的纯品融合蛋白,SDS-PAGE鉴定两融合蛋白表达正确。说明PlnEPlnF基因片段编码的多肽能在原核细胞中正确表达,为进一步研究该细菌素的生物活性奠定了基础。

关键词: 植物乳酸杆菌; 细菌素; 原核表达; 融合蛋白

Abstract: In order to obtain high-yield pure plantaricin, and research on biological veterinary, plantaricin PlnE and PlnF were synthesised by Invitrogen. The aim genes were inserted into a bacterial expression vector pGEX-4T-1 to construct two prokaryotic expression vectors of pGEX-4T-E and pGEX-4T-F. After IPTG induction and express optimization, the fusion proteins were purified by affinity chromatography and detected by SDS-PAGE. The results showed that expression vectors were constructed correctly, more than 30% pure fusion proteins were obtained, which indicated peptides PlnE and PlnF could be accurately expressed in prokaryote cells, and lay a foundation for further researches on their bioactivity.

Key words: plantaricin; bacteriocin; heterologous expression; fusion protein

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