CONSTRUCTION OF A FAMILY OF MYCOBACTERIUM ESCHERICHIA COLI SHUTTLE VECTORS DERIVED FROM PAL5000 AND PACYC184 - THEIR USE FOR CLONING AN ANTIBIOTIC-RESISTANCE GENE FROM MYCOBACTERIUM-FORTUITUM/

The pSUM vectors, a family of Mycobacterium/Escherichia coli shuttle c loning vectors derived from the pSU series of vectors are described. T hey all contain the pACYC184 origin of replication enabling them to re plicate in E. coli, the mycobacterial pAL5000 origin of replication an d a kanamycin-resistance (Km(R)) gene which allows selection in both E . coli and mycobacteria. They carry a multiple cloning site (MCS) to f acilitate cloning and the lacZ alpha reporter gene for screening inser ts in E. coli. A library of total DNA from a strain of Mycobacterium f ortuitum was constructed with one of these vectors, pSUM36, and a myco bacterial gentamicin-resistance (Gm(R)) gene was cloned by selecting f or Gm(R) in Mycobacterium smegmatis.