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. 1987 Dec;169(12):5423–5428. doi: 10.1128/jb.169.12.5423-5428.1987

Facile autoplast generation and transformation in Bacillus thuringiensis subsp. kurstaki.

I T Crawford 1, K D Greis 1, L Parks 1, U N Streips 1
PMCID: PMC213967  PMID: 3680166

Abstract

We describe a method for maximizing the rate of conversion of Bacillus thuringiensis subsp. kurstaki vegetative cells to osmotically fragile forms in the absence of exogenously added enzymes. Optimal generation of autoplasts occurred in 50 mM sodium acetate buffer (pH 7.0) at 37 degrees C with 10% (wt/vol) polyethylene glycol as an osmotic stabilizer. The maximum autolytic rate resulted in a conversion of greater than 90% of bacilli to spherical autoplasts in 6 min. Autoplasts regained bacillary morphology upon plating on DM3-G regeneration medium, with reversion frequencies ranging from 1.2 x 10(-1) to 5.3 x 10(-3). The autoplasts could efficiently take up exogenously added plasmid DNA. The presence of plasmids was verified by Southern hybridization analysis.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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