scholarly journals Expression and purification of recombinant coat protein of sugarcane mosaic virus from Indonesian isolate as an antigen for antibody production

2019 ◽  
Vol 24 (1) ◽  
pp. 57
Author(s):  
Natalia Tri Astuti ◽  
Nurmalasari Darsono ◽  
Suvia Widyaningrum ◽  
Widhi Dyah Sawitri ◽  
Sri Puji Astuti ◽  
...  
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Polyclonal Antibodies ◽  
Molecular Study ◽  
Insoluble Fraction ◽  
Sugarcane Mosaic Virus ◽  
Gene Encoding ◽  
Iptg Induction ◽  
Sds Page ◽  
Saccharum Sp

Sugarcane mosaic virus (SCMV, genus Potyvirus, family Potyviridae) is a prominent pathogen of sugarcane (Saccharum sp. hybrids). It can cause losses in susceptible varieties, in crop as well as sugar production, economically. Although it has been studied in major sugar-producing countries, research on the definement of SCMV from Indonesian isolates based on molecular study has been very limited. This study aimed to obtain a proper recombinant antigens emanating from coat protein of SCMV from Indonesian isolate in order to produce polyclonal antibodies that cann be used for immunodiagnosis assays in a subsequent study. A gene-encoding coat protein of SCMV (CP-SCMV) was amplified using RT-PCR and cloned into vector pJET1.2. The cDNA was inserted into 6X His-tag expression plasmid of pET28a(+) and over-expressed in Escherichia coli BL21(DE3) to produce a recombinant protein. The highest expression was found in 0.1M IPTG induction media for 5 h at 37oC. SDS-PAGE analysis clarified that the recombinant CP-SCMV remained as an insoluble fraction. Purifications was carried out by the affinity Ni-NTA resin, followed by electroelution to obtain a highly purified protein. To meet the quality requirements of a proper antigen, the highly purified protein was concentrated. A molecular weight of the rCP-SCMV (approximately 40 kDa) was clearly observed by 10% SDS-PAGE at the concentration of 16.184 mg/mL. 

scholarly journals Triticum mosaic virus: A New Virus Isolated from Wheat in Kansas

Plant Disease ◽  
2008 ◽  
Vol 92 (5) ◽  
pp. 808-817 ◽  
Author(s):  
Dallas L. Seifers ◽  
T. J. Martin ◽  
Tom L. Harvey ◽  
John P. Fellers ◽  
James P. Stack ◽  
...  
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Polyclonal Antibodies ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cesium Chloride ◽  
High Plains ◽  
Flight Mass Spectrometry ◽  
Sds Page ◽  
Systemic Symptoms

In 2006, a mechanically-transmissible and previously uncharacterized virus was isolated in Kansas from wheat plants with mosaic symptoms. The physiochemical properties of the virus were examined by purification on cesium chloride density gradients, electron microscopy, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), sequencing of the nucleotides and amino acids of the coat protein, and immunological reactivity. Purified preparations contained flexuous, rod-shaped particles that resembled potyviruses. The coat protein was estimated from SDS-PAGE to have a mass of approximately 35 kDa. Its amino acid sequence, as deduced from DNA sequencing of cloned, reverse-transcribed viral RNA and separately determined by time-of-flight mass spectrometry, was most closely related (49% similarity) to Sugarcane streak mosaic virus, a member of the Tritimovirus genus of the family Potyviridae. The virus gave strong positive reactions during enzyme-linked immunosorbent assays using polyclonal antibodies raised against purified preparations of the cognate virus but gave consistent negative reactions against antibodies to Wheat streak mosaic virus (WSMV), other wheat potyviruses, and the High Plains virus. When the virus was inoculated on the WSMV-resistant wheat cv. RonL, systemic symptoms appeared and plant growth was diminished significantly in contrast with WSMV-inoculated RonL. Taken together, the data support consideration of this virus as a new potyvirus, and the name Triticum mosaic virus (TriMV) is proposed.

scholarly journals Characteristics of gene encoding coat protein isolated from soybean mosaic virus

2014 ◽  
Vol 36 (1se) ◽  
Author(s):  
Lo Thi Mai Thu ◽  
Hoang Ha ◽  
Le Van Son ◽  
Chu Hoang Ha ◽  
Chu Hoang Mau
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Soybean Mosaic Virus ◽  
Gene Encoding

Transformation of Wheat With the Gene Encoding the Coat Protein of Barley Yellow Mosaic Virus

1996 ◽  
Vol 23 (4) ◽  
pp. 429 ◽  
Author(s):  
S Karunaratne ◽  
A Sohn ◽  
A Mouradov ◽  
J Scott ◽  
HH Steinbiss ◽  
...  
Keyword(s):  
Gene Expression ◽  
Coat Protein ◽  
Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Bar Gene ◽  
Gene Encoding ◽  
Barley Yellow Mosaic Virus ◽  
Virus Coat Protein ◽  
Wheat Embryos ◽  
Virus Coat

Wheat plants (Triticum aestivum cv. Hartog) were stably transformed with the bar gene and the gene encoding the barley yellow mosaic virus coat protein. Cultured immature wheat embryos were bombarded with tungsten particles coated with the pEmuPAT-cp construct. Fifteen regenerating 'PPT- resistant' plants were selected on medium containing phosphinothricin. Of these, 11 plants had both the bar and cp genes integrated into the wheat genome and two plants had only the bar gene. Transmission of the two genes to progeny of two independent plants was confirmed. The barley yellow mosaic virus coat protein was detected in both the parent and progeny plants; however, bar gene expression occurred only in the parent plants.

Full sequence of the coat protein gene is required for the induction of pathogen-derived resistance against sugarcane mosaic virus in transgenic sugarcane

2018 ◽  
Vol 45 (6) ◽  
pp. 2749-2758 ◽  
Author(s):  
Retnosari Apriasti ◽  
Suvia Widyaningrum ◽  
Weny N. Hidayati ◽  
Widhi D. Sawitri ◽  
Nurmalasari Darsono ◽  
...  
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Sugarcane Mosaic Virus ◽  
Full Sequence ◽  
Transgenic Sugarcane ◽  
Pathogen Derived Resistance

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.

Expression, purification, and use as an antigen of recombinant sugarcane mosaic virus coat protein

1995 ◽  
Vol 140 (10) ◽  
pp. 1817-1831 ◽  
Author(s):  
G. R. Smith ◽  
R. Ford ◽  
J. D. Bryant ◽  
R. L. Gambley ◽  
T. K. McGhie ◽  
...  
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Sugarcane Mosaic Virus ◽  
Virus Coat Protein ◽  
Virus Coat
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