Cloning and sequencing 5' and 3' SipA gene of Salmonella enteritidis in E.coli
Subject Areas : Aquatic ProductsGazizeh Shirazi 1 , Abbas Doosti 2
1 - کارشناسی ارشد ژنتیک، مرکز تحقیقات بیوتکنولوژی، دانشگاه آزاد اسلامی واحد شهرکرد
2 - استادیار، مرکز تحقیقات بیوتکنولوژی، دانشگاه آزاد اسلامی واحد شهرکرد
Keywords: Escherichia coli, Cloning, Salmonella enteritidis, SipA gene,
Abstract :
Introduction: Salmonella infections in many domesticated animals, wildlife and humans cause disease all of the world. The ability to enter and survive in host cells is a prerequisite for pathogenicity species of Salmonella. Salmonella invasion protein is an important virulence factors into host cells by bacteria transmitted. In this study we showed cloning the 5 'and 3' gene SipA Salmonella and was performed in E. coli. Materials & Methods: In this study, by PCR and specific primers sequences of 5' and 3' SipA gene was amplified correctly. DNA fragment was cloned by T/A cloning technique in pGEM T-easy vector and transformed into E. coli. Results: Cloning of SipA gene was confirmed by PCR. The results of next step showed that the SipA gene was successfully cloned in E. coli. Final confirmation of construct was done by XbaI, BglII, SacI, XhoI restriction enzymes. Conclusion: According to the results that, insertion of antibiotic-resistant genes between 5' and 3' regions of the SipA gene of Salmonella entritidis due to gene deletion in the bacteria. The construct's gene can be used as a target for gene vaccines against Salmonella entritidis in the future.
