Molecular cloning of S1 glycoprotein gene of infectious bronchitis virus (IBV) serotype 793/B in secretory Pichia pastoris vector
In vitro protein expression is an important method of obtaining large amounts of viral proteins to investigate their biological properties. The S1 glycoprotein of infectious bronchitis virus, due to its effective immune-dominant role is an appropriate candidate for production of recombinant vaccine against infectious bronchitis disease. In this study, the S1 gene fragment of infectious bronchitis virus strain793/B was amplified using reverse transcriptase-polymerase chain reaction (RT-PCR) and purified. It
was then cloned into pPICZαA a secretory expression vector of Pichia pastoris. The insertion was proved by PCR analysis and isolation of gene from construct by restriction enzymes and finally, it was sequenced. After the expression of S1 gene in P. pastoris expression system, it was found that it could be used in the production of recombinant vaccines against infectious bronchitis disease.
Keywords: Infectious bronchitis, S1 glycoprotein, cloning, Pichia pastoris