Transformation of vegetative cells of Bacillus anthracis with plasmid DNA

Journal of General Microbiology
C P Quinn, B N Dancer

Abstract

Methods have been developed for chemical transformation and electro-transformation (electroporation) of vegetative cells of Bacillus anthracis with supercoiled plasmid DNA. Chemical transformation was dependent on incubation in Tris/HCl with osmotic support and transformation with plasmid DNA was effected by treatment with polyethylene glycol 3350. Maximum transformation frequencies were 3.8 x 10(-5) transformant c.f.u. per viable c.f.u. (1 x 10(3) c.f.u. per micrograms DNA). Optimal frequencies were pH dependent and were affected by growth-medium composition. Transformation was not observed with linear or multimeric plasmid DNA. Electro-transformation of B. anthracis using high field intensity electroporation was dependent on the composition of both the growth medium and the electroporation buffer. Maximum electro-transformation frequencies were 5.3 x 10(-4) c.f.u. per viable c.f.u. (2.6 x 10(4) c.f.u. per micrograms DNA). The use of early exponential phase cells was critical to both procedures and the maximum efficiency (c.f.u. per micrograms DNA) of each system was strain dependent under the conditions described.

Citations

May 28, 2009·Applied Microbiology and Biotechnology·Naoto Yoshida, Misa Sato
Apr 2, 2008·Antimicrobial Agents and Chemotherapy·G C KedarRobert J Haselbeck
Jun 24, 2003·Infection and Immunity·Marcus B Jones, Martin J Blaser
Feb 24, 2006·Infection and Immunity·Brian K Janes, Scott Stibitz
Feb 3, 2009·Journal of Bacteriology·Jared D HeffronDavid L Popham
Mar 15, 2006·Plasmid·Christina TamDominique Missiakas
Dec 20, 2011·Gene·Ramakrishnan Sitaraman, Stephen H Leppla
Nov 28, 2012·Microbial Pathogenesis·Jared D HeffronSusan L Welkos
Oct 7, 2004·The Journal of Biological Chemistry·Xudong LiangNicholas S Duesbery
Mar 29, 2000·Protein Expression and Purification·S Park, S H Leppla

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