OPRL CLONING OF PSEUDOMONAS AERUGINOSA IN ESCHERICHIA COLI BL21 AFTER BIOINFORMATICS VERIFICATION AS A CANDIDATE FOR VACCINE.

Background and Aim:Pseudomonas aeruginosa is a Gram-negative bacterium that is considered to be one of the most important bacterial pathogens responsible for serious opportunistic infections among cystic fibrosis (CF) and immunocompromized patients. Perhaps, choosing the right vaccine can solve the problem. So, we suggest other membrane protein L (OprL).Methods:Primer design, PCR (Polymerase Chain Reaction) and cloning for oprL in the plasmid pET 28a done after bioinformatics studies. Then, its transfer to Escherichia coli BL21 was performed. Finally, it was approved by universal primer PCR and enzymatic digestion methods.Results:The bioinformatics studies shown protein conservation. Also, it was strong antigenic and immunogenic properties for OprL by IEDB Analysis and vaxijen software respectively. In the laboratory experiments, the oprL was cloned in Escherichia coli BL21. Finally, the approval tests were positive.Conclusion:This protein is suitable for evaluating animal experiments.

Keywords:

Vaccine , Pseudomonas aeruginosa , Clonning , oprL