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枯草芽孢杆菌绿色荧光蛋白高效表达载体的构建
引用本文:刘 伟,林志伟,陈美霞,魏日凤,谢联辉. 枯草芽孢杆菌绿色荧光蛋白高效表达载体的构建[J]. 热带作物学报, 2012, 33(3): 467-471
作者姓名:刘 伟  林志伟  陈美霞  魏日凤  谢联辉
作者单位:1. 福建农林大学植物病毒研究所 生物农药与化学生物学教育部重点实验室 福建福州350002
2. 宁德师范学院生物工程系,福建宁德,352100
3. 福建农林大学园艺学院,福建福州,350002
基金项目:农业部948项目(No. 2009-Z11);福建省高校新世纪人才资助项目;福建省专家服务团项目;福建省农科教结合资助项目。
摘    要:利用绿色荧光蛋白基因gfpmut3标记枯草芽孢杆菌(Bacillus subtilis)的启动子P43;构建1个枯草芽孢杆菌-大肠杆菌穿梭表达载体pP43GFP。结果表明:P43与gfpmut3构成了融合基因,可调控gfpmut3在枯草芽孢杆菌和大肠杆菌中的表达;荧光检测发现,P43在枯草芽孢杆菌和大肠杆菌中具有极强的启动基因表达能力;质粒稳定性测定表明,连续稀释培养55 h后,质粒的稳定性为85%。

关 键 词:绿色荧光蛋白基因;枯草芽孢杆菌;穿梭载体

Construction of High-Level Expression of Green Fluorescent Protein Vector in Bacillus subtilis
LIU Wei,LIN Zhiwei,CHEN Meixi,WEI Rifeng and XIE Lianhui. Construction of High-Level Expression of Green Fluorescent Protein Vector in Bacillus subtilis[J]. Chinese Journal of Tropical Crops, 2012, 33(3): 467-471
Authors:LIU Wei  LIN Zhiwei  CHEN Meixi  WEI Rifeng  XIE Lianhui
Affiliation:Institute of Plant Virology, Fujian Agriculture and Forestry University, Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education;Institute of Plant Virology, Fujian Agriculture and Forestry University, Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education;Department Biological Engineering of Ningde Normal University;College of Horticulture, Fujian Agriculture And Forestry University;Institute of Plant Virology, Fujian Agriculture and Forestry University, Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education
Abstract:The P43 promoter from Bacillus was chosen to drive the expression of gfp-mut3 and was cloned into B.subtilis-Escherichia coli shuttle plasmid,the recombinant of plasmid pP43GFP obtained was transformed into recipient strains of E.coli DH5α and B.subtilis.The result showed that gfpmut3 gene driven by P43 could express not only in B.subtilis but also in DH5α and that the stability of pP43GFP was 85% after the continuous diluted cultivation for 55 hours.
Keywords:Green fluorescent protein(GFP)gene  Bacillus subtilis  Shuttle vector
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