Open AccessCloning and expression of the origin of replication of mycobacteriophage D29 in Mycobacterium smegmatis
Author(s) -
Lazraq R.,
HoussainiIraqui M.,
ClavelSérès S.,
David H.L.
Publication year - 1991
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1991.tb04646.x
Subject(s) - mycobacterium smegmatis , shuttle vector , cloning (programming) , biology , genetics , vector (molecular biology) , kanamycin , molecular cloning , puc19 , plasmid , microbiology and biotechnology , recombinant dna , gene , computational biology , gene expression , mycobacterium tuberculosis , medicine , tuberculosis , pathology , computer science , programming language
Summary Libraries of mycobacteriophage D29 genes were obtained by transforming Escherichia coli with constructs derived from pUC19. The genomic libraries were used to transform Mycobacterium smegmatis MC 2 155, and one of the vectors designated pRM64 was found to stably replicate in the mycobacterial recipients. The pRM64 vector contained a 2.65‐kb fragment that was used as a probe and was then located on the physical map of D29. Vectors containing this fragment replicated stably in M. smegmatis for at least 144 generations in medium without the selective agent used (kanamycin); vectors not containing the fragment did not replicate in the recipient M. smegmatis . This is the first report showing that the cloning of the OriR of a mycobacteriophage allowed the stable replication of shuttle vectors for mycobacterial genetics.