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Virologica Sinica, 20 (5) : 515-518, 2005
Expression of M1 protein of Avian Influenza Virus and Analysis of Its Immunoreactivity
A pair of primers were designed based on M1 gene sequence of known H5N1 Avian influenza virus.M1 gene was cloned from total RNA,extracted from tissue of H5N1 subtype virus inoculated embryo by reverse transcriptase-polymerase chain reaction using high proofreading polymerase(Pyobest~TM DNA Polymerase),and expressed using Invitrogen champion~TM pET directional TOPO expression system.Recombinant protein containing polyhistidine(6xHis) tag in N-terminal about 29.8kDa in size,wac obtained and purified.Its immunoreactivity was analyzed by Western blot,ELISA and blocking ELISA using MAb and positive serum.The results showed recombinant M1 protein can bind to MAb and positive serum with specificity,and this binding reactivity can be blocked by natural viral antigen.This indicates that the recombinant M1 protein possesses good immunoreactivity.
  Published online: 20 Oct 2005
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