Construction of shuttle vectors for genetic manipulation and molecular analysis of mycobacteria

Gene
S JainA K Tyagi

Abstract

Two novel shuttle vectors for mycobacteria are described which have been derived from the expression system pSD5 developed in our laboratory. Plasmid pSD5B is a promoter-selection vector containing a promoterless lacZ gene and allows the identification of mycobacterial promoters by the blue colour of the colonies on solid media containing XGal. Moreover, the chronological order of appearance of blue colonies and intensity of colour provide a qualitative index of transcriptional strengths of the cloned promoters. Plasmid pSD5C has been designed to construct mycobacterial genomic libraries and express the cloned DNA inserts as fusion proteins with maltose binding protein in mycobacteria. Libraries in pSD5C provide feasibility for their screening with either DNA probes or specific antisera for identifying the genes of interest and for isolation of specific genetic loci by complementation of Escherichia coli and mycobacterial mutants. These vectors combine the ease of working in E. coli with the advantage of directly propagating them in mycobacteria without further manipulations. Finally, we demonstrate that these vectors function efficiently both in fast growing Mycobacterium smegmatis and slow growing mycobacteria including Mycob...Continue Reading

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Citations

Apr 30, 2013·Tuberculosis·Nisha RathorMridula Bose
Aug 31, 2010·The Journal of Biological Chemistry·Michael J MortonMichael J Caplan
Apr 27, 2000·Current Opinion in Pulmonary Medicine·T M Doherty, P Andersen
Apr 7, 2012·Archives of Microbiology·Sukhendu Mandal, Sujoy K Das Gupta
Feb 22, 2011·Tuberculosis·Anil K TyagiVijaya Satchidanandam
Oct 27, 2015·Nucleic Acids Research·Wareed AhmedValakunja Nagaraja
Oct 27, 2009·Archives of Biochemistry and Biophysics·Pabitra K ParuaPradeep Parrack
Nov 18, 2003·Molecular Microbiology·Amy I KimGraham F Hatfull
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Jul 14, 2010·FEMS Microbiology Letters·Tara Rao, Peter A Lund
Oct 22, 2003·FEMS Microbiology Letters·Amit SinghAnil K Tyagi
May 22, 2016·Biochemical and Biophysical Research Communications·Soumitra GhoshValakunja Nagaraja
Nov 1, 2005·Tuberculosis·Ramandeep SinghAnil K Tyagi
Apr 21, 2019·Applied and Environmental Microbiology·Abhishek Anil Dubey, Vikas Jain
Dec 21, 2000·Molecular Microbiology·S Jain, G F Hatfull

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