TY - JOUR
T1 - Development of a new host vector system in mycobacteria
AU - Goto, Yoshitaka
AU - Taniguchi, Hatsumi
AU - Udou, Takezo
AU - Mizuguchi, Yasuo
AU - Tokunaga, Tohru
N1 - Funding Information:
We thank Mr. T. Yamazaki( NationalI nstitute of Health) and Ms. M. Ogawa and K. Mizue (Universityo f Occupationaal nd Environmental Health) for their technicalh elp. This investigation was supported by the grants from the SasakawaM emorialHealth Foundationa nd the Japan Health SciencesF oundation.
PY - 1991/10/15
Y1 - 1991/10/15
N2 - The hybrid plasmid pYT72/pYT92 constructed from an Escherichia coli plasmid pACYC177 and mycobacterial plasmid pMSC262 isolated from Mycobacterium scroflaceum strain W262 transformed both E. coli and BCG. Phage-sensitive mutants S-10 and S-20 isolated from BCG Tokyo strain showed higher frequency of transformation than the wild-type strain. Frequency of transformation was dependent on age of the culture and the electroporation condition used. Several deletion mutants were generated from pYT72/92 to determine the minimum region for the replication in the mycobacteria. A 2.3-kb fragment of pMSC262 was found to contain an essential region. Using this fragment and pACYC177, a small shuttle vector pYT937 containing two drug-resistance markers, kanamycin- and ampicillin-resistance, was constructed. pYT937 contains AatII, BamHI, BbvII, GsuI, HincII, PstI, ScaI and XbaI cloning sites.
AB - The hybrid plasmid pYT72/pYT92 constructed from an Escherichia coli plasmid pACYC177 and mycobacterial plasmid pMSC262 isolated from Mycobacterium scroflaceum strain W262 transformed both E. coli and BCG. Phage-sensitive mutants S-10 and S-20 isolated from BCG Tokyo strain showed higher frequency of transformation than the wild-type strain. Frequency of transformation was dependent on age of the culture and the electroporation condition used. Several deletion mutants were generated from pYT72/92 to determine the minimum region for the replication in the mycobacteria. A 2.3-kb fragment of pMSC262 was found to contain an essential region. Using this fragment and pACYC177, a small shuttle vector pYT937 containing two drug-resistance markers, kanamycin- and ampicillin-resistance, was constructed. pYT937 contains AatII, BamHI, BbvII, GsuI, HincII, PstI, ScaI and XbaI cloning sites.
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U2 - 10.1111/j.1574-6968.1991.tb04477.x
DO - 10.1111/j.1574-6968.1991.tb04477.x
M3 - Article
C2 - 1769534
AN - SCOPUS:0026425713
VL - 83
SP - 277
EP - 282
JO - FEMS Microbiology Letters
JF - FEMS Microbiology Letters
SN - 0378-1097
IS - 3
ER -