A constitutive expression vector pHY300-Flgfp was constructed to test the function of promoter F1 subcloned from a rice epiphyte Bacillus brevis strain DX01. The DX01 cells harboring plasmid pHY300-F1gfp were detected to produce bright green fluorescence. Subsequently, the gfp-tagged B. brevis strain was released into the soil and its survival was investigated by PCR and the detection of green fluorescence. The spatial location of in situ gfp-tagged bacterial cells on the root surface of rice seedlings was visualized. All these results indicated that green fluorescent protein is an ideal molecular marker for the detection of the activities of promoter F1, and it is also a reliable probe to monitor specific B. brevis bacteria in the environment.

Department of Plant Science School of Agriculture and Biology Shanghai Jiaotong University Shanghai 201101 PR China

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FEBS Lett. 1994 Mar 21;341(2-3):277-80 PubMed

Science. 1994 Feb 11;263(5148):802-5 PubMed

J Microbiol Methods. 1999 Apr;35(3):187-99 PubMed

Curr Biol. 1996 Feb 1;6(2):178-82 PubMed

Curr Microbiol. 2001 Oct;43(4):244-8 PubMed

Gene. 1999 Feb 4;227(1):101-10 PubMed

Gene. 1992 Feb 15;111(2):229-33 PubMed

J Appl Bacteriol. 1995 Feb;78(2):97-108 PubMed

Appl Environ Microbiol. 1997 Nov;63(11):4543-51 PubMed

J Microbiol Methods. 2000 Apr;40(2):135-45 PubMed

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