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Appl. Environ. Microbiol., Sep 1995, 3256-3260, Vol 61, No. 9
Copyright © 1995, American Society for Microbiology

Use of Electroporation To Generate a Thiobacillus neapolitanus Carboxysome Mutant

RS English, S Jin and JM Shively
Department of Biological Sciences, Clemson University, Clemson, South Carolina 29634-1903

Two cloning vectors designed for use in Escherichia coli and the thiobacilli were constructed by combining a Thiobacillus intermedius plasmid replicon with a multicloning site, lacZ(prm1), and either a kanamycin or a streptomycin resistance gene. Conditions necessary for the introduction of DNA into T. intermedius and T. neapolitanus via electroporation were examined and optimized. By using optimal electroporation conditions, the gene encoding a carboxysome shell protein, csoS1A, was insertionally inactivated in T. neapolitanus. The mutant showed a reduced number of carboxysomes and an increased level of CO(inf2) necessary for growth.

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