Obtaining of the p17 recombinant protein of human immunodeficiency subtype A virus |
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Authors: | R I Al-Shekhadat I V Dukhovlinov A I Kobatov N A Klimov A P Kozlov |
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Institution: | 1.State Research Institute for Highly Pure Biopreparations,Federal Medicobiological Agency and Biomedical Center Research Organization,St. Petersburg,Russia |
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Abstract: | A simple and efficient method for expression in Escherichia coli cells and purification of a recombinant matrix protein, p17, of human immunodeficiency type I virus has been described. HIV-1
subtype A DNA sequence encoding p17 was obtained by amplification of the viral gag gene segment and cloned into an expression vector under the control of T7Lac promoter. The conditions for cell growth and
induction of p17 synthesis by lactose and its further purification by metal chelate chromatography were optimized. p17 preparations
with 97% purity were obtained; the yield of the protein of 28 mg per 1l of culture was achieved. The obtained protein was
capable of binding antibodies from blood serum of a HIV-infected patient during immunoblotting. |
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