Genetic manipulation of Desulfovibrio.

Publisher Summary This chapter describes genetic manipulation of Desulfovibrio. In several cases, it has been observed that expression in Escherichia coli ( E. coli ) of genes for Desulfovibrio metal proteins results in the accumulation of apoproteins, sometimes as insoluble complexes without the metal (clusters) incorporated, because of the lack of proper processing of such proteins. Introduction of such genes on expression vectors in Desulfovibrio results in a considerable (over)production of the proteins in their native form, allowing purification in high amounts and facilitating biochemical analysis. To prevent accumulation of the inhibitory sulfide, sulfate might be replaced by nitrate for strains that are able to use nitrate as a terminal electron acceptor. Plating of Desulfovibrio cells in a soft-agar overlay is recommended to prevent swarming of the cells. IncQ plasmids are not self-transmissable and require trans-acting functions for mobilization. These trans-acting functions (tra genes) are normally derived from the IncP plasmid RP4 and are either incorporated into the genome of the E. coli donor or provided on helper plasmids.

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