Abstract:

In this paper, the broad-host vector pBB1MCS-5 was used to construct an expression vector for flippase
recombination enzyme (FLP). The expression vector was transformed into Gluconobacter suboxydans, and the positive
transformants were screened. The results of validation using a two-step RT-PCR method showed that flp was transcribed and
expressed in the host cell. The pBBR1MCS-psldh-flp recombinant plasmid was transformed into tetracyclic-resistant JGDH－
mutant strains, and PCR verification showed that the sites with FTR resistance marker had been effectively removed.
Recombination FLP expression in Gluconobacter suboxydans provides a method that circularly uses the selectable marker
gene to knock out or replace multiple locus genes.

Key words: Gluconobacter suboxydans, FLP/FRT, site-specific recombination, promoter, gene knock-out