[Construction of a GFP broad-host-range promoter-probe vector and its application]

The 0.9 kb DNA fragment was obtained by PCR and using a pair of special designed primers from the green fluorescent protein gene(gfp). It Was then cloned into an expression vector pET-11C and named as pHN115. A promoferless gfp vector with SD fragment was obtained from pHN115 and subcloned into SK(+), pIJ 2925 and finally into pTR102. A broad-hast-range and visible promoter-probe vector, pHN127 was constructed and used successfully for the identification of constitutive and inducible promoters from Sinorhizobium fredii HN01.