Open AccessExpression of green fluorescent protein in Bacillus brevis under the control of a novel constitutive promoter F1 and insertion mutagenesis of F1 in Escherichia coli DH5α
Author(s) -
Chen Yunpeng,
Yan Jian,
Yang Mingjie,
Wang Jingwen,
Shen Daleng
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00797-3
Subject(s) - escherichia coli , mutagenesis , promoter , biology , green fluorescent protein , site directed mutagenesis , microbiology and biotechnology , bacillus (shape) , fluorescence , genetics , gene , mutation , gene expression , physics , mutant , quantum mechanics
The constitutive expression vector pHY300‐F1gfp was constructed to test the function of a promoter, F1, cloned from the rice epiphyte Bacillus brevis strain DX01. The DX01 cells harboring the plasmid pHY300‐F1gfp were shown to produce bright green fluorescence. The results were confirmed by Western blot analysis and fluorescence‐activated cell sorting. Expression of the F1 promoter was constitutive. To improve the activity of F1, insertion mutagenesis of F1 based on in vitro transposition reaction was performed. Seven mutants with enhanced transcription activity in Escherichia coli DH5α were obtained. The enhanced promoters showed similar high activities in B. brevis strain DX01.