کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8359649 | 1542313 | 2016 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Expression, purification and characterization of a full-length recombinant HIV-1 Vpu from inclusion bodies
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کلمات کلیدی
βTrCPBST-2VPUTMBPMSFIPTGDTTHRPTBS3,3′,5,5′-tetramethylbenzidine - 3،3 '، 5،5'-تترامیلیل بنزیدینisopropyl β-D-1-thiogalactopyranoside - ایزوپروپیل β-D-1-thiogalactopyranosideRefolding - بازپرداختOver-expression - بیش از بیانTris-buffered saline - تریس بافر شورEnzyme-linked immunosorbent assay - تست الیزاELISA - تست الیزاSolubilization - حلال سازیdithiothreitol - دیتیوتریتولendoplasmic reticulum - شبکه آندوپلاسمی Phenylmethylsulfonylfluoride - فنیل متیل سولفونیل فلوئوریدHIV-1 - ویروس اچ آی وی نوع یکPurification - پاکسازیHorseradish peroxidase - پراکسیداز هوررادیش
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Vpu is one of four accessory proteins encoded by human immunodeficiency virus type I (HIV-1). Vpu modulates the expression of several cellular restriction factors within the HIV-1 infected cell including CD4, CD74, the bone marrow stromal antigen 2 (BST-2) and NK-T-and-B antigen. The interaction of HIV-1 Vpu with these proteins interferes with the innate immune response directed against HIV-1; thereby promoting viral persistence. The involvement of HIV-1 Vpu in manipulating the cellular environment in ways that favor viral replication makes it an attractive target for anti-HIV drug intervention. This paper describes the over-expression and purification of a soluble HIV-1 Vpu from inclusion bodies by ion-exchange chromatography, allowing production of 6Â mg of highly purified protein (>95% purity) per 10Â mg of pelleted cells obtained from 1Â L of bacterial culture. Far-UV circular dichroism showed that the recombinant protein is folded and retained its secondary structure. Moreover, using ELISA, known HIV-1 Vpu binding partners, BST-2 and CD74, showed that the refolded purified protein is functional or at least assumes a conformation that is capable of binding these putative binding partners. To our knowledge, this is the first report of the purification and successful solubilization of full-length, wild-type HIV-1 Vpu from inclusion bodies in Escherichia coli.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 128, December 2016, Pages 109-114
Journal: Protein Expression and Purification - Volume 128, December 2016, Pages 109-114
نویسندگان
Zikhona Njengele, Ronel Kleynhans, Yasien Sayed, Salerwe Mosebi,