期刊论文

中文

合成生物学

2096-8280

2019

5

71-75

10.3969/j.issn.1674-0319.2019.05.009

湖南省研究生科研创新项目(CX2017B388) 湖南省自然科学基金(2016JJ4038) 湖南省教育厅科研项目(15K062).

本校为第一机构

根据绿色荧光蛋白（green fluorescent protein,GFP）的特性,将GFP作为目的基因,与穿梭载体p HY300PLK结合,构建一个芽孢杆菌WB600的蛋白表达体系,为在枯草芽孢杆菌中表达其他蛋白提供了一定的技术和理论支持。本文结合p HY300PLK的克隆位点,设计gfp基因的引物,PCR扩增后回收的gfp基因片段进行双酶切,与相同双酶切的pHY300PLK连接获得pHY300PLK-GFP,通过电转化将pHY300PLK-GFP转化至芽孢杆菌WB600中并检测。获得转化子WB1,检测到转化子在荧光显微镜下显示绿色荧光,且后续测序检测表明重组质粒构建成功。

According to the characteristics of green fluorescent protein(GFP),GFP was used as the target gene combined with the shuttle vector pHY300PLK to construct a protein expression system basing on Bacillus sp.WB600.The primers of gfp gene were designed according to the cloning site of pHY300PLK.The fragment of gfp gene was recovered by PCR and double-digested.The pHY300PLK-GFP was obtained by connecting the fragment with double-digested pHY300PLK, transforming pHY300PLK-GFP into Bacillus sp.WB600 by electrical transformation and detected.The transformant WB1 was obtained,and the transformant showe...