Cloning and expression of the filamentous bacteriophage Pf1 major coat protein gene in Escherichia coli

1987 ◽  
Vol 195 (4) ◽  
pp. 873-884 ◽  
Author(s):  
David H. Rowitch ◽  
Richard N. Perham
Keyword(s):  
Escherichia Coli ◽  
Coat Protein ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Cloning And Expression ◽  
Major Coat Protein ◽  
Filamentous Bacteriophage

Molecular Cloning and Expression of the Coat Protein Gene of Plum Pox Virus El-Amar Strain in E.coli

2004 ◽  
Vol 1 (1) ◽  
pp. 2-2
Author(s):  
Abou El- Nasr . ◽  
M. A, Dougdoug . ◽  
K. A., Hayam S. Abde . ◽  
Rehab A. Dawoud .
Keyword(s):  
Coat Protein ◽  
Molecular Cloning ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Cloning And Expression ◽  
Plum Pox Virus

scholarly journals Expression of Grapevine leafroll-associated virus 3 coat protein gene in Escherichia coli and production of polyclonal antibodies

2007 ◽  
Vol 32 (6) ◽  
pp. 496-500 ◽  
Author(s):  
Thor V.M. Fajardo ◽  
Danielle R. Barros ◽  
Osmar Nickel ◽  
Gilmar B. Kuhn ◽  
F. Murilo Zerbini
Keyword(s):  
Escherichia Coli ◽  
Coat Protein ◽  
Western Blot ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Polyclonal Antibodies ◽  
Yield And Quality ◽  
Sds Page ◽  
Grapevine Leaves

Grapevine leafroll-associated virus 3 (GLRaV-3), the main viral species of the grapevine leafroll complex, causes yield and quality reduction in grapes (Vitis spp.). The coat protein gene was RT-PCR-amplified from total RNA extracted from infected grapevine leaves and the amplified fragment was cloned and completely sequenced. The fragment was subsequently subcloned into the pRSET-C expression vector. The recombinant plasmid was used to transform Escherichia coli BL21:DE3 and express the capsid protein. The coat protein, fused to a 6 His-tag, was purified by affinity chromatography using an Ni-NTA resin. The identity of the purified protein was confirmed by SDS-PAGE and Western blot. The in vitro-expressed protein was quantified and used for rabbit immunizations. The antiserum was shown to be sensitive and specific for the detection of GLRaV-3 in grapevine extracts in Western blot and DAS-ELISA assays, with no unspecific or heterologous reactions against other non-serologically related viruses being observed.

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