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Abstract Objective The goal of the current study was to express the pp85 gene of human herpesvirus 7 (HHV-7) and to prepare anti-pp85 polyclonal antibodies. Methods The pp85 gene of HHV-7 was amplified using PCR and this amplified gene was cloned into a prokaryotic expression vector and named as PGEX-6P-1+pp85b. The recombinant plasmid was then transformed into Escherichia coli Rosetta. The accuracy of the inserted gene and the specificity of the proteins were verified by restriction enzyme digestion,SDS-PAGE,and Western blot. The expressed pp85 fusion protein was then purified using affinity chromatography and was used to immunize rabbits in order to obtain the anti-pp85 antibodies. The specificity of these antibodies was verified using immunofluorescence. Results The purity of the recombinant pp85 was above 90% and the titer of anti-pp85 antibodies was 1∶102 400. As an indication of the specificity of the antibodies produced,this polyclonal antibody was shown to react with HHV-7 infected cells but failed to react with human herpesvirus 6 (HHV-6) infected cells as shown by IFA.Conclusion In the current study we were able to obtain a highly purified form of the HHV-7 pp85 fusion protein and the corresponding high-titer polyclonal antibodies were produced in rabbit.
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Received: 01 January 1900
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Corresponding Authors:
YAO Kun
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