Abstract
Objective To purify and crystallize two kinds of H5N1 vires hemagglutinin proteins, H5 I151F and H5 I151F + A134V + E186D, from the cellular membrane. Methods Recombinant vaccinia viruses were massive propagated, hemagglutinin (HA) proteins were extracted from cellular membrane with detergent. HA proteins were concentrated with continuous sucrose gradient ultracentrifugation and detected by Western Blotting. HA proteins were cleavaged with Bromelain, and purified with Ion-exchange chromatography. The protein puri-ty was detected by SDS-PAGE electrophoresis and staining. HA proteins were crystallized by sitting-drop vapour diffusion. Results Hemag-glutinin proteins, H5 I151F and H5 I151F + A134V + E186D, were extensively purified. And the crystal of H5 I151F was obtained. Con-clusion For the first time, the highly purified H5 I151F membrane protein crystal was obtained, which provided the basis for further stud-ying the mechanisms of human to human transmission caused by avian influenza viruses. Key words: Influenza,avian; Influenza A virus,avian/IP; Hemagglutinin glycoproteins,influenza virus/IP
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