Application of flagellar hook protein FlgE of reorganized pseudomonas aeruginosa
A technology of flagellar hook protein and Pseudomonas aeruginosa, which can be applied in the direction of medical preparations containing active ingredients, antibacterial drugs, drug combinations, etc., and can solve problems affecting the extension and function of flagellar body
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[0025] Recombinant expression and purification of FlgE (small-scale preparation)
[0026] According to the FlgE coding gene sequence of Pseudomonas aeruginosa, the upstream and downstream primers for amplifying the full-length FlgE coding sequence with NdeI and HindIII restriction sites were designed and synthesized. The above primers were used to amplify the extracted Pseudomonas aeruginosa genomic DNA. The PCR program was: denaturation at 94°C for 45 seconds, annealing at 58°C for 1 minute, extension at 72°C for 1 minute, and 25 cycles; After separation and purification by agarose gel electrophoresis, it was digested with NdeI and HindIII and recovered, and ligated with the expression vector PET24a that had undergone the same digestion; the ligated product was transformed into E. 50 μg / mL) on LB agar plate, select Kan-resistant clones after overnight culture, extract the plasmid after shaking the bacteria for amplification, and carry out enzyme digestion identification. Afte...
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