Physiological factors involved in the transformation of Mycobacterium smegmatis

M. V. Norgard, T. Imaeda

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Transfer of streptomycin resistance and changes from methionine and leucine auxotrophy to prototrophy were achieved in M. smegmatis by transformation. Recipient cells were more resistant to mitomycin C and methyl methanesulfonate treatments than were wild-type cells. A high level of calcium ions was essential for transformation, especially during DNA adsorption, whereas the presence of magnesium ions and the exposure of recipient cells to mild doses of UV light enhanced recombination frequencies. Transformants were not isolated when recipient cell-DNA mixtures were first treated with deoxyribonuclease. Recipient cells at various stages of growth showed similar transformabilities. Transformation was successful only when recipient cells were incubated on rich agar medium after mixture with DNA. Exposure of recipient cells to Pronase before treatment with donor DNA did not affect transformation, suggesting the absence of a protein competence factor. Throughout the present experiments, cotransformation frequencies were very low and unselected marker segregation patterns were independent, indicating that the methionine, leucine, and streptomycin markers are not closely linked in M. smegmatis.

Original languageEnglish (US)
Pages (from-to)1254-1262
Number of pages9
JournalJournal of bacteriology
Volume133
Issue number3
StatePublished - Jan 1 1978

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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