to construct a transcriptional fusion of the ram gene cluster promoter to egfp the PCL product generated using the primers pramS1 and pramS2 was digested with BglII and NdeI and cloned between the BglII and NdeI and NdeI sites of pHL117. this generated pHL171,which contains the major promoter of the ram gene cluster, Piram. pKF59 isan integrative plasmid containing a divIVAegfp translational fusion gene. It was digested with StuI and cloned at the SmaI site of pOJ260. this generated pHL175 ,which has an RP4-derived origin of transfer(oriT) region to facilitate E.coli-Streptomyces conjugation.