The Prokaryotic Expression and Identification of 33K Gene of Fowl Adeno-virus Group Ⅰ
The non-structural protein 33K of fowl adenovirus group Ⅰ is important for virus infection.The purpose of this study is to construct prokaryotic expression vector pGEX-33K for the diagnosis antigen.According to the sequences of FAVⅠ 33K gene available in GenBank,a pair of primers was designed.The target sequence was amplified by PCR,and was ligated to prokaryotic vector pGEX-4T-1 to construct recombinant expression plasmid pGEX-33K.The recombinant plasmid was transformed into Escherichia coli DH5α,and then induced and expressed by IPTG induction.The pGEX-33K recombinant protein was analyzed by SDS-PAGE and Western Blot.The result showed that pGEX-33K was mainly expressed in the form of soluble proteins and the molecular weight of the fusion protein was about 36.7 KD.The optimal concentration of IPTG and the time was 0.75 mmol/L and 4 h,respectively.Furthermore,the result of the Western Blot demonstrated that the pGEX-33K could be specifically recognized by positive serum to FAVⅠ,which might be used as the coating antigen to develop a differential diagnosis kit against fowl adenovirus group Ⅰ.
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