Construction and application of plasmid pZCY11 for analyzing gene functions and expressions in Myxococcus

Abstract:

Abstract: [Objective] To construct a plasmid for analyzing gene functions and expressions and to study the MXAN1334 gene in Myxococcus xanthus with the plasmid. [Methods] We constructed the plasmid vector pZCY11, amplified MXAN1334 gene fragment from M. xanthus DK1622 by PCR, and inserted the fragment into a site upstream of lacZ, resulting in the recombinant plasmid pZCY13. The plasmid pZCY13 was transformed by electroporation to DK1622, producing a mutant ZC16-18 (?MXAN1334). [Results] The plasmid pZCY11 carried the resistance gene aph as the selectable marker, the replication origin of OriR6K and promoterless reporter gene lacZ. We examined the swarm expansions of ZC16-18 on CTT hard and soft agar, and the result indicated that MXAN1334 gene was probably involved in gliding motility in M. xanthus. In addition, β-galactosidase activity of ZC16-18 was detected by X-gal assay and the blue color developed was used to mark the colony growth. Time of colour showed that MXAN1334 gene was expressed in the early stage in M. xanthus. [Conclusion] The plasmid vector pZCY11 made it more convenient for the study on functions and the expressions of target gene in M. xanthus.