Preserved antigenicity of HIV-1 p24 produced and purified in high yields from plants inoculated with a tobacco mosaic virus (TMV)-derived vector

Journal of Virological Methods
D M Pérez-FilgueiraT J Morris

Abstract

Production of structural proteins from foot-and-mouth disease virus (FMDV) and bovine herpes virus (BHV-1) in Nicotiana benthamiana through the use of a tobacco mosaic virus-based vector (TMV-30B) has been reported previously. The development of the TMV-30B-HISc vector, a new version that adds a C-terminal histidine (His) sequence to the foreign protein expressed is described. Coding sequences from the FMDV VPl protein and the core protein, p24, from a clade C HIV-1 isolate from Zambia were cloned into the new vector and infective RNAs were generated for each construct to inoculate N. benthamiana plants. His-tagged proteins were purified from inoculated leaves using immobilized metal affinity chromatography (IMAC) as detected by Coomassie blue staining and proteins were further characterized in Western blot assays using a commercial anti-6xHis mAb and specific polyclonal antisera for each protein. While yields obtained for the VPl-His protein after purification were similar to those in crude extracts obtained with the previous TMV-VPl vector, p24-His yields were 10-15 times higher than those of VPl-His. Twenty-five grams of TMV-p24-HISc inoculated leaves were processed to obtain 2.5 mg of isolated p24-His and the recombinant pr...Continue Reading

References

Apr 18, 1992·Lancet·J J LefrèreM H Elghouzzi
Dec 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·H S MasonC J Arntzen
Oct 1, 1990·AIDS Research and Human Retroviruses·L S EhrlichC A Carter
Nov 5, 1997·Protein Expression and Purification·J NilssonP A Nygren
Jan 20, 1999·Proceedings of the National Academy of Sciences of the United States of America·A A McCormickR Levy
Feb 7, 2001·Current Opinion in Biotechnology·A M Walmsley, C J Arntzen
Nov 4, 2000·Nature Biotechnology·G GiddingsA Carter
Apr 3, 2001·Science·D Paydarfar, W J Schwartz
Dec 18, 2001·International Journal of Cancer. Journal International Du Cancer·H MantinaC Wood
Mar 6, 2002·Molecular Biotechnology·G Gary ZhangK Andrew White
Jun 6, 2002·The Plant Journal : for Cell and Molecular Biology·Rachel L TothSean Chapman
Jul 31, 2002·Annual Review of Phytopathology·Gregory P PogueWayne P Fitzmaurice
Feb 25, 2003·The Journal of Infectious Diseases·Brad P BrayfieldCharles Wood
May 7, 2003·Current Opinion in Biotechnology·Amanda M Walmsley, Charles J Arntzen
Jan 1, 1997·Annual Review of Phytopathology·J JohnsonG Lomonossoff
Jul 22, 2008·Biotechnology and Bioengineering·A R KusnadiJ A Howard

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Citations

Jun 25, 2008·BMC Biotechnology·Ann MeyersEdward P Rybicki
Aug 3, 2010·Expert Review of Vaccines·Nunzia ScottiFranco Maria Buonaguro
May 26, 2009·Biotechnology Advances·Sharon M-H Shih, Pauline M Doran
Jan 10, 2009·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Ingrid LindhSören Andersson
Nov 20, 2012·Research in Veterinary Science·Margherita FestaAlfonso Zecconi
Feb 15, 2020·Current Opinion in Biotechnology·Jocelyne Tremouillaux-GuillerAbdullah Makhzoum

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