Development of a broad-host-range sacB-based vector for unmarked allelic exchange.

ABSTRACT:

Background

Although genome sequences are available for an ever-increasing number of bacterial species, the availability of facile genetic tools for physiological analysis have generally lagged substantially behind traditional genetic models.

Results

Here I describe the development of an improved, broad-host-range "in-out" allelic exchange vector, pCM433, which permits the generation of clean, marker-free genetic manipulations. Wild-type and mutant alleles were reciprocally exchanged at three loci in Methylobacterium extorquens AM1 in order to demonstrate the utility of pCM433.

Conclusion

The broad-host-range vector for marker-free allelic exchange described here, pCM433, has the advantages of a high copy, general Escherichia coli replicon for easy cloning, an IncP ori

SUBMITTER: Marx CJ 

PROVIDER: S-EPMC2518277 | biostudies-literature | 2008 Feb

REPOSITORIES: biostudies-literature