USE OF ELECTROPORATION TO GENERATE A THIOBACILLUS-NEAPOLITANUS CARBOXYSOME MUTANT

Two cloning vectors designed for use in Escherichia coli and the thiob acilli were constructed by combining a Thiobacillus intermedius plasmi d replicon with a multicloning site, lacZ', and either a kanamycin or a streptomycin resistance gene. Conditions necessary for the introduct ion of DNA into T. intermedius and T. neapolitanus via electroporation were examined and optimized. By using optimal electroporation conditi ons, the gene encoding a carboxysome shell protein, esoS1A, was insert ionally inactivated in T. neapolitanus. The mutant showed a reduced nu mber of carboxysomes and an increased level of CO2 necessary for growt h.