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Virologica Sinica, 21 (1) : 6-10, 2006
Research Article
Expression of Herpes Simplex Virus 1 Glycoprotein D in Pichia pastoris
Chinese Academy of Medical Sciences /Peking Union Medical college
Institute of Medical Biology Kunming China
The extracellular portion of glycoprotein D gene(gDt),encoding 1-314 amino acids,was amplified from extracted HSV-1 DNA and cloned into expression vector pPIC9K.After electroporation transformation of Pichia pastoris GS115 and selection with G418,a strain of P.pastoris with high yield of secreted recombinant gD was obtained.The yield of recombinant gD was 250mg/liter in shake flask cultures.The recombinant protein could be identified with specific monoclonal antibody 1-I-9 using ELISA and Western blotting assays. Recombinant gD was purified to almost homogeneity by
Q-Sepharose ion exchange, Chelating Sepharose immobilized metal ion affinity and Sephacryl S-200
gel filtration chromatographies. Purified gD expressed in P. pastoris elicited high level of specific
antibodies in BALB/c mice, indicating recombinant gD were characteristic high immunogenicity and
capable of inducing significant titers of specific antibodies.
  Published online: 20 Jan 2006
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