Analysis of the replication region of a mycobacterial plasmid, pMSC262

M. Qin, Hatsumi Taniguchi, Y. Mizuguchi

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Abstract

We determined the nucleotide sequence of a DNA fragment which contains the replication region of pMSC262, a Mycobacterium scrofalaceum plasmid used to construct the Mycobacterium-Escherichia coli shuttle vector. The complete sequence of the fragment contained 2,504 bp with an overall G+C content of 69.8%. By deletion analysis, we found that the minimum length required for plasmid replication in M. bovis BCG was about 1.6 kb. Within this region, several open reading frames (ORFs) and a putative replication origin (ori) were identified by computer analysis. One of the ORFs, ORF2, which encodes a putative 28.9-kDa basic protein with characteristics of DNA-binding proteins, appeared to be involved in replication of the plasmid in BCG. By separation of ORF2 and the putative ori region, it was revealed that the relative locations of ORF2 and the putative ori region are likely important for replication in BCG. No DNA or amino acid homologies were found between this replication region and that of pAL5000, another mycobacterial plasmid used for vector plasmid construction. In addition, we found that this replicon did not lead to replication in E. coli and was compatible in BCG with pAL5000- derived vector plasmid pYUB75 (R. G. Barletta, D. D. Kim, S. B. Snapper, B. R. Bloom, and W. R. Jacobs, J., J. Gen. Microbiol. 138:23-30, 1992).

Original languageEnglish
Pages (from-to)419-425
Number of pages7
JournalJournal of bacteriology
Volume176
Issue number2
DOIs
Publication statusPublished - Jan 1 1994
Externally publishedYes

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Plasmids
Mycobacterium bovis
Mycobacterium
Open Reading Frames
Escherichia coli
Genetic Vectors
Replicon
Replication Origin
DNA
Base Composition
DNA-Binding Proteins
Amino Acids
Proteins

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Cite this

Analysis of the replication region of a mycobacterial plasmid, pMSC262. / Qin, M.; Taniguchi, Hatsumi; Mizuguchi, Y.

In: Journal of bacteriology, Vol. 176, No. 2, 01.01.1994, p. 419-425.

Research output: Contribution to journalArticle

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