Immunodiagnosis of groundnut and watermelon bud necrosis viruses using polyclonal antiserum to recombinant nucleocapsid protein of Groundnut bud necrosis virus.
In vitro gene expression strategy was used for the production of polyclonal antiserum to the nucleocapsid protein (NP) of Groundnut bud necrosis virus (GBNV). The GBNV NP gene from cowpea isolate was cloned into 6x His-tagged UA cloning vector and expressed in Escherichia coli [M15] cells. The fusion protein was detected in insoluble fraction and was purified by using Ni-NTA agarose resin. The purified 6x His-fusion protein ( approximately 32 kDa) was used for immunisation to produce a high titre polyclonal antiserum. The antiserum to the NP of GBNV at 1:4000 dilution detected successfully natural infection of GBNV and Watermelon bud necrosis virus in a wide range of cucurbitaceous, leguminous and solanaceous hosts from different locations.
Jain
R K
RK
Unit of Plant Virology, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi 110012, India. rakeshjain56@yahoo.co.in
Pandey
Amar N
AN
Krishnareddy
M
M
Mandal
Bikash
B
D000383:Agriculture / Q000379:methods; D000818:Animals; D010367:Arachis / Q000821:virology*; D029773:Citrullus / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007106:Immune Sera; D019590:Nucleocapsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D010935:Plant Diseases / Q000821:virology*; D011817:Rabbits; D017865:Tospovirus / Q000737:chemistry / Q000302:isolation & purification*
D007106:Immune Sera; D019590:Nucleocapsid Proteins