Obtaining of the p17 recombinant protein of human immunodeficiency subtype A virus
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 in...
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| Veröffentlicht in: | Applied biochemistry and microbiology 2011-12, Vol.47 (8), p.730-736 |
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| container_title | Applied biochemistry and microbiology |
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| creator | Al-Shekhadat, R. I Dukhovlinov, I. V Kobatov, A. I Klimov, N. A Kozlov, A. P |
| description | 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. |
| doi_str_mv | 10.1134/S0003683811080023 |
| format | Article |
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I ; Dukhovlinov, I. V ; Kobatov, A. I ; Klimov, N. A ; Kozlov, A. P</creator><creatorcontrib>Al-Shekhadat, R. I ; Dukhovlinov, I. V ; Kobatov, A. I ; Klimov, N. A ; Kozlov, A. P</creatorcontrib><description>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.</description><identifier>ISSN: 0003-6838</identifier><identifier>EISSN: 1608-3024</identifier><identifier>DOI: 10.1134/S0003683811080023</identifier><language>eng</language><publisher>Dordrecht: Springer-Verlag</publisher><subject>Antibodies ; Biochemistry ; Biomedical and Life Sciences ; Blood ; blood serum ; Cell culture ; cell growth ; Cellular biology ; Chelates ; Chromatography ; E coli ; Escherichia coli ; Expression vectors ; Gag protein ; genes ; HIV ; Human immunodeficiency virus ; Human immunodeficiency virus 1 ; humans ; Immunoblotting ; immunosuppression (physiological) ; Lactose ; Life Sciences ; matrix protein ; Medical Microbiology ; Metals ; Microbiology ; Nucleotide sequence ; nucleotide sequences ; Promoters ; protein purification ; Proteins ; Virology ; viruses</subject><ispartof>Applied biochemistry and microbiology, 2011-12, Vol.47 (8), p.730-736</ispartof><rights>Pleiades Publishing, Ltd. 2011</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c366t-eff5dc4e478fab7cc548a3489b8a0e351a9bb456271450c41f87d671cfb0655e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1134/S0003683811080023$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1134/S0003683811080023$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>315,782,786,27933,27934,41497,42566,51328</link.rule.ids></links><search><creatorcontrib>Al-Shekhadat, R. 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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. 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| subjects | Antibodies Biochemistry Biomedical and Life Sciences Blood blood serum Cell culture cell growth Cellular biology Chelates Chromatography E coli Escherichia coli Expression vectors Gag protein genes HIV Human immunodeficiency virus Human immunodeficiency virus 1 humans Immunoblotting immunosuppression (physiological) Lactose Life Sciences matrix protein Medical Microbiology Metals Microbiology Nucleotide sequence nucleotide sequences Promoters protein purification Proteins Virology viruses |
| title | Obtaining of the p17 recombinant protein of human immunodeficiency subtype A virus |
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