Use of Green Fluorescent Protein Gene to Label Bacillus subtilis Strains

The gfp sequence was amplified from pAD123 by PCR.pAXc-gfp was constructed by ligation of the gfp PCR product and pAXc digested by BamHI.pAXc-gfp was linearized by Pst I and transformed into B.subtilis B411.The linearized plasmid should be integrated at the lacA locus on B.subtilis B411 chromosome DNA.The recombinants were screened by chloramphenicol resistance(5 μg/mL) phenotype.Recombinant named B412 could express GFP stably without antibiotic pressure.