Salmonella typhi whip Mao Suji for fljB- z66 Cloning expression and polyclonal antibody preparation.doc

 PAGE \* MERGEFORMAT 15 Salmonella typhi whip Mao Suji for fljB: z66 Cloning expression and polyclonal antibody preparation Study: Health Xiu Zhang Fanggao Yu Lin Huang Xinxiang High-XU of Chee Soon Juan River [Abstract] Objective: Cloning and Expression of H: z66 positive Salmonella typhi whip Mao Suji for fljB: z66, and purified the corresponding polyclonal antibodies. Methods: Using PCR technology from the H: z66 positive Salmonella typhi genomic DNA obtained by whip Mao Suji fljB: z66, cloned into the expression vector pET 28a (+) and E. coli JM109 in on to express; fljB: z66 expressed product was purified by Ni was used as antigen to prepare polyclonal antibodies in rabbits. Results : The PCR and sequence analysis confirmed that due to Salmonella typhi whip Mao Suji fljB: z66 expression vector was successfully imported pET 28a (+), and Escherichia coli JM109 was highly expressed as a rabbit anti-antigen was successfully prepared z66 polyclonal antibodies. Conclusion: cloning and expression of the H: z66 positive Salmonella typhi whip Mao Suji for fljB: z66, and prepared the appropriate polyclonal antibodies for future in-depth study H: z66 positive Salmonella typhi, one-way phase the mechanisms underlying transformation. [Keywords:] Salmonella typhi; fljB: z66; polyclonal antibody [Abstract] Objective: To clone and express the flagellin gene fljB: z66 of H: z66 positive Salmonella enterica serover Typhi, and prepare the anti z66 polyclonal antibodies.Methods: Specific primers were designed to amplify the flagellin gene fljB: z66 from the chromosome DNA of H: z66 positive Salmonella enterica serover Typhi GIFU10007.The amplicon was inserted into the expression vector pET 28a (+), which was then transferred to E.coli JM109 to be expressed.The recombinant protein FljB His6 was purified with Ni TED packed column and was used as immunogen to prepare the polyclonal antibody.Results: PCR and sequencing analysis demonstrated that the flagellin gene flj