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Archives of Microbiology

, Volume 136, Issue 4, pp 275–280 | Cite as

Transfection of Mycobacterium smegmatis SN2 with mycobacteriophage I3 DNA

  • S. Sadashiva Karnik
  • K. P. Gopinathan
Original Papers

Abstract

Mycobacterium smegmatis SN2 does not exhibit natural competence for the uptake of phage I3 DNA. Competence can artificially be induced by treatment with glycine or CaCl2, and the combination of both is even more effective. The efficiency of transfection can be improved by inclusion of protamine sulphate and heterologous RNA in the system. From 32P DNA uptake studies the major barrier for the entry of DNA has been found to be the complex cell wall. The efficiency of transfection calculated on the basis of fraction of DNA which has entered the cell is comparable to that of other bacterial systems. The phage development takes a longer time (7 h for one cycle) after transfection, as compared to infection (4 h).

Key words

Transfection Mycobacterium smegmatis Mycobacteriophage I3 Calcium sensitization Glycine sensitization Protamine sulfate Exogenous RNA DNA uptake 

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Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • S. Sadashiva Karnik
    • 1
  • K. P. Gopinathan
    • 1
  1. 1.Microbiology and Cell Biology LaboratoryIndian Institute of ScienceBangaloreIndia

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