2013•Progress in Veterinary MedicineRequires access
Prokaryotic Expression of chIRF7 Protein and Preparation of Polyclonal Antibody
Ming Liao
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Abstract
The chIRF7 gene was amplified by RT-PCR and inserted into pET30a vector to construct recombinant plasmid pET30a-chIRF7.Then,the pET30a-chIRF7 was transformed into BL21(DE3) and induced with IPTG,the SDS-PAGE results indicated that the recombinant protein rchIRF7 was about 60 ku.The purified recombinant protein was used to immunize mice to prepare polyclonal antibody,the titer of the polyclonal antibody was above 1∶51 200 detected with indirect ELISA,the IFA test results indicated that the polyclonal antibody could recognize chIRF7 protein expressed in CEF cells stimulated by AIV specifically,it revealed that the chIRF7 recombinant protein had high immunogenicity and the prepared polyclonal antibody had high titer and specificity.