植物乳杆菌细菌素基因plnEF的克隆与异源表达

目的:构建目的基因plnEF原核表达系统,诱导工程菌BL21-p ET28a-PL-plnEF表达目的融合蛋白并纯化,以期获得大量纯度较高的植物乳杆菌PlnEF细菌素,开发新的食品防腐剂。方法:构建含plnEF基因的重组质粒,将其转入大肠杆菌BL21中,经过IPTG诱导大量表达目标蛋白,融合蛋白PlnEF经纯化后进行分子量大小、抑菌活性等的检测。结果:重组质粒pET28a-PL-plnEF在大肠杆菌BL21中成功表达,合成PlnEF蛋白,其分子量为15.6 k Da,且该融合蛋白对大肠杆菌JM109具有良好的抑菌活性。结论:plnEF基因片段能够在原核细胞中正确表达且具有活性,本文为进一步研究开发该细菌素作为生物防腐剂奠定了基础。

Clone and Heterologly Expression of Bacteriocin Gene pln EF of Lactobacillus plantarum

Objective: The prokaryotic expression system of the target gene plnEF was constructed,and the engineered strain BL21-pET28 a-PL-plnEF was induced to express and purify the target fusion protein,so as to obtain a large amount of the higher purity Lactobacillus plantarum PlnEF bacteriocin and develop a new food preservative.Method: The recombinant plasmid containing plnEF gene was constructed and transferred into E. coli BL21.The target protein was expressed by IPTG,and the fusion protein PlnEF was purified to detect the molecular weight and antibacterial activity. Result: The recombinant plasmid pET28 a-PL-plnEF was successfully expressed in E.coli BL21,and the PlnEF protein was synthesized with a molecular weight of 15.6 kDa. The fusion protein had good antibacterial activity against Escherichia coli JM109. Conclusion: The plnEF gene fragment could be correctly expressed and active in prokaryotic cells,which would lay a foundation for further research and development of the bacteriocin as a biological preservative.