Polyclonal Antibodies to the Coat Protein of Carnation etched ring virus Expressed in Bacterial System: Production and Use in Immunodiagnosis^†

Abstract

Carnation etched ring virus (CERV)

Preparation of Polyclonal Antibodies to Grapevine fanleaf Virus Coat Protein Expressed in Escherichia coli

“…Like other virus systems such as Faba bean necrotic yellows virus (Kumari et al, 2001), sugarcane yellow leaf virus (Korimbocus et al, 2002), Potato mop-top virus (Cerovska et al, 2003(Cerovska et al, , 2006 and Groundnut bud necrosis virus (Jain et al, 2005) where tagged viral CP was expressed in insoluble fraction, the fusion GFLV CP was detected in the insoluble fraction (pellet) from the transformed E. coli. Expression of recombinant fusion protein in the soluble fraction has also been reported elsewhere (Hema et al, 2003;Hourani and Abou-Jawdah, 2003;Lee et al, 2008;Raikhy et al, 2007), but we detected only a small amount of the fusion protein in the soluble fraction.…”

“…The use of purified expressed fusion protein as the antigen gives consistent results in terms of quantity and quality of the antigen (Raikhy et al, 2007). Thus, the recombinant viral CPs expressed in bacterial cells have great potentials as the alternative sources of antigens for raising specific antibodies to plant viruses.…”

“…Antibodies against recombinant proteins of a number of plant viruses have been prepared successfully (Iracheta-Cardenas et al, 2008;Lee and Chang, 2008;Raikhy et al, 2007;Jain et al, 2005;Abou-Jawdah et al, 2004;Nickel et al, 2004;Cerovska et al, 2003;Hourani and Abou-Jawdah, 2003;Korimbocus et al, 2002;Kumari et al, 2001). By the use of different IPTG concentrations it was concluded that the optimal expression was achieved, when transformed E. coli Rosetta cells was induced with 1 mM IPTG at OD = 0.5 and grown at 37°C for 4 h. Different IPTG concentrations including 1 mM (Bragard et al, 2000;Kadkhodayan et al, 2000;Saini and Vrati, 2003;Thomas and Baneyx, 1996) and 0.4 mM (Jacob and Usha, 2002) have been reported as the optimal concentration with various genes of interest for expression of Cardamom mosaic virus CP and 0.1 mM (Petrzik et al, 2001) for that of Prunus necrotic Ring Spot virus (PNRSV) CP in E. coli.…”

Diagnosis of a new variant of soybean yellow mottle mosaic virus with extended host-range in India

“…The recombinant approach is advantageous as it does not encounter the problems associated with the mixture of closely related viruses, purification and maintenance of live virus culture. This strategy has been successfully used for a number of plant viruses from different genera however not for carmoviruses [4,9,13,16]. To our knowledge, this is the first report of development of immunodiagnosis based on antiserum against bacterial expressed recombinant CP of any carmoviruses in general and SYMMV in particular.…”

Molecular characterization of Indian isolate of peanut mottle virus and immunodiagnosis using bacterial expressed core capsid protein

“…Similar results of expression of recombinant His-tagged CP in the insoluble fraction have been reported for several plant viruses, pelargonium zonate spot virus [27], faba bean necrotic yellows virus [15], sugarcane yellow leaf virus [13], potato mop-top virus [5,6] and groundnut bud necrosis virus [12]. However, for some plant viruses such as sugarcane streak mosaic virus, cucurbit yellow stunting disorder virus, cymbidium mosaic virus, the His-tagged viral CP was expressed in soluble fraction [10,11,16,23]. The PAb against core CP successfully detected the expressed protein as well as the virus in infected peanut leaf samples.…”