{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,17]],"date-time":"2025-09-17T16:50:17Z","timestamp":1758127817505},"reference-count":45,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2017,6,13]],"date-time":"2017-06-13T00:00:00Z","timestamp":1497312000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech and App Biochem"],"published-print":{"date-parts":[[2018,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Virion infectivity factor (Vif) is a 23 kDa protein that protects HIV\u20101 from deamination of its proviral DNA by APOBEC3G. The active form of Vif is a multimer that interacts simultaneously with CBF\u2010beta, the elongin B and C subunits, Cullin 5, and APOBEC3G to form a ubiquitin ligase complex targeting the latter for degradation. Vif clearly represents an attractive target for developing novel antiviral drugs for the therapy of HIV\/AIDS, and this goal requires a source of well folded, readily available protein. For that purpose, we have cloned<jats:italic>Vif<\/jats:italic>in the pET28a expression vector, expressing the resulting His\u2010tagged recombinant protein in the BL21(DE3)<jats:italic>Escherichia coli<\/jats:italic>strain. After lysis, Vif was solubilized from the insoluble fraction with 6 M guanidinium chloride and purified by denaturing immobilized\u2010metal affinity chromatography, refolding the protein afterwards by dialysis. The use of 2\u2010(<jats:italic>N<\/jats:italic>\u2010morpholino)ethanesulfonic acid buffer at pH 6.2 and the presence of EDTA improved Vif refolding yields by reducing the formation of insoluble aggregates. The purified protein was bound by two monoclonal antibodies against sequential and conformational epitopes located at the C and N terminus, respectively.<\/jats:p>","DOI":"10.1002\/bab.1557","type":"journal-article","created":{"date-parts":[[2017,2,9]],"date-time":"2017-02-09T04:53:05Z","timestamp":1486615985000},"page":"195-202","source":"Crossref","is-referenced-by-count":1,"title":["An improved method for purification and refolding of recombinant HIV Vif expressed in<i>Escherichia coli<\/i>"],"prefix":"10.1002","volume":"65","author":[{"given":"Carlos A.","family":"Duarte","sequence":"first","affiliation":[{"name":"Physical\u2010Chemistry Division Bioinformatics Department Center for Genetic Engineering and Biotechnology La Habana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mickel","family":"Palomino","sequence":"additional","affiliation":[{"name":"Physical\u2010Chemistry Division Bioinformatics Department Center for Genetic Engineering and Biotechnology La Habana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2017,6,13]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature00939"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2003.10.034"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1386-6532(02)00113-0"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature01709"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nm946"},{"key":"e_1_2_7_7_1","doi-asserted-by":"crossref","first-page":"2867","DOI":"10.1101\/gad.1250204","volume":"18","author":"Yu Y.","year":"2004","journal-title":"Genes Dev."},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2007.07.029"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature10693"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/nm945"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1089591"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.1496"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/cmdc.201200079"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.06957-11"},{"key":"e_1_2_7_15_1","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1186\/1743-422X-11-122","volume":"11","author":"Matsui M.","year":"2014","journal-title":"Virol. 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