Prokaryotic Expression of African Swine Fever F334L Gene Expressing Protein and Preparation of Its Polyclonal Antibody
The recombinant expression plasmid pCold-Ⅰ-ASFV-F334L was constructed by cloning the F334L gene encoding one of non-structural protein of African swine fever virus(ASFV)into the prokaryotic expression vector pCold-Ⅰ.The pCold-Ⅰ-ASFV-F334L was transformed into competent cells,and the prokaryotic expression was induced by 1 mmol/L IPTG.Through SDS-PAGE and Western blot experiments,the results indicated that the F334L gene was well expressed.The purified F334L gene-encoded protein was used to immunize mice,resulting in the production of polyclonal antibodies in the serum.The generated antibodies were used to detect the recombinant porcine reproductive and respiratory syndrome virus expressing ASFV F334L gene-encoded protein(rPRRSV-F334L)constructed and rescued by the research team.The experimental results demonstrated that the polyclonal antibodies exhibited good reactivity and specificity,proving that the ASFV F334L gene can be stably expressed in the PRRSV live vector,thereby providing material for the development of ASFV live vector vaccines.
African Swine Fever Virusviral live vectorF334L generecombinant virus
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