产品介绍

pGMB151 is a derivative of the suicide plasmid pKNG101 and includes oriR6K, RP4, sacBR, insB, and strAB

with the bla gene from plasmid pBR322 cloned at the Xba I site. It is a suicide plasmid because of its requirement for the pir protein, which is supplied by kpir in certain strains of Escherichia coli, such as SM10.

To improve the efficiency of allelic exchange for genome genetic modification,a novel pGMB152 vector was constructed with thelacZYA operon incorporated into pGMB151 to provideLacZ-based blue–white selection. The LacZ gene, a well-known reporter gene expressing b-galactosidase, can provide blue–white selection to make the existence of molecular cloning visible  and could be used as a biomarker for the suicide plasmid. Bacteria colonies withpGMB152 will show as blue and those without pGMB152 will show as white; therefore, bacterial colonies with different colors could be distinguished easily during the different processes of genome genetic modification. The new plasmid was used to knock out the hsdM gene [9] in Salmonellapullorum to construct a candidate-attenuated Salmonella vaccine, to repair pseudogene flhA by knockin and pseudogene flgk by site-directed mutagenesis.

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