Isolation and characterization of the replicon of a Thiobacillus intermedius plasmid.

Published

Journal Article

The replicon of a cryptic Thiobacillus intermedius plasmid (pTiK12) has been isolated and sequenced. Functional analysis of deletion subclones in Escherichia coli localized the replicon to a 3.5-kb region of DNA. Sequencing of this region identified a 30-bp A-T-rich potential stem-loop structure. In addition, an 11-bp direct repeat, an 11-bp inverted repeat, and a 16-bp inverted repeat were observed at the stem-loop structure. Also found in the replicon was a series of four tandem direct repeats consisting of a perfectly conserved 8-bp core. A region near the stem-loop structure is involved in the regulation of plasmid copy number. Deletion subclones lacking this region have increased copy numbers, indicating a negative regulatory role. An open reading frame capable of encoding a 320-amino-acid protein was found near the stem-loop structure. The putative amino acid sequence shares significant similarity with the two Rep proteins from the ColE2 and ColE3 replicons. Replication of the T. intermedius replicon is dependent upon DNA polymerase I. The isolation and examination of the T. intermedius plasmid replicon are initial steps toward the establishment of a genetic system in T. intermedius.

Full Text

Duke Authors

Cited Authors

  • English, RS; Lorbach, SC; Huffman, KM; Shively, JM

Published Date

  • January 1995

Published In

Volume / Issue

  • 33 / 1

Start / End Page

  • 1 - 6

PubMed ID

  • 7753904

Pubmed Central ID

  • 7753904

International Standard Serial Number (ISSN)

  • 0147-619X

Digital Object Identifier (DOI)

  • 10.1006/plas.1995.1001

Language

  • eng

Conference Location

  • United States