Fusion expression,detection and application of HHV-8 antigen / 重庆医学

ABSTRACT

Objective To construct the prokaryotic expression plasmid of HHV-8 fusion antigen for diagnosis of HHV-8 infec-tion .Methods The combined fragment ORF59 ,ORF65 and K8 .1 by fusion PCR was integrated into pQE-80L and transfected into E .coli DH5α.Fusion protein was induced to express by IPTG .SDS-PAGE and Western blot were employed to detect the fusion protein .Fusion protein was used to detect serum of blood donors .Results The combined plasmid pQE-80L-ORF59-ORF65-K8 .1 was constructed successfully after verifying by restriction enzyme digestion and sequencing .The fusion protein was about 24 KD and could be specific combined with HHV-8 positive serum .The fusion protein had the same result to detect HHV-8 with the HHV-8 ELISA kit .Conclusion Fusion protein we construct can be used as diagnosis antigen to detect HHV-8 of blood donors and common people .