Development of a new host vector system in mycobacteria

Yoshitaka Goto, Hatsumi Taniguchi, Takezo Udou, Yasuo Mizuguchi, Tohru Tokunaga

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)277-282
Number of pages6
JournalFEMS microbiology letters
Volume83
Issue number3
DOIs
Publication statusPublished - Oct 15 1991

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology
  • Genetics

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