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Volume 148, Issue 7

Comparative evaluation of as a surrogate cloning host for use in the study of mycobacterial genetics Free

Abstract

represents an alternative mycobacterial cloning host that has been largely overlooked to date. The main reason for this may be the reported non-transformability of this species, specifically the so-called Stanford strain (NCTC 11659), with expression vectors that use kanamycin resistance as a selection method. However, this strain can be transformed using hygromycin resistance as an alternative selectable phenotype. The present study has shown that in contrast to previous reports, (ATCC 15483) is capable of being transformed with a range of vectors encoding kanamycin resistance as the selectable marker. Thereafter, the expression of the reporter gene in , BCG and mc155 was evaluated using a range of characterized mycobacterial promoter sequences (, , AN, and ) cloned in the same promoter probe vector. In general, the promoters showed similar levels of activity in the three species, demonstrating that existing expression systems can readily be employed with (ATCC 15483). This was further confirmed by the observation that was capable of stable, expression of recombinant S1 subunit of pertussis toxin at levels equivalent to those obtained with BCG and . Analysis of structural and functional stability of a range of vectors demonstrated that the incidence of instability noted for was lower than that recorded for . Taken together, the results indicate that is an additional cloning host which may prove useful for specific aspects of mycobacterial biology and provide increased flexibility to the field of recombinant protein technology for mycobacteria.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): expression , mycobacteria , shuttle vectors and stability
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2002-07-01
2024-04-20
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Comparative evaluation of Mycobacterium vaccae as a surrogate cloning host for use in the study of mycobacterial genetics
Microbiology 148, 1999 (2002); https://doi.org/10.1099/00221287-148-7-1999
/content/journal/micro/10.1099/00221287-148-7-1999
/content/journal/micro/10.1099/00221287-148-7-1999
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