Over expression of Recombinant CP3 Protein of Rice Tungro Spherical Virus in Prokaryotic Expression System
Currently, serological based diagnosis of rice tungro disease (RTD) is being limited by the availability of antisera. This study aims to produce recombinant protein from the coat protein (CP)3 of rice tungro spherical virus (RTSV) as a potential antigen in producing high titre antibodies for use...
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| Main Authors: | , , |
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| Format: | E-Article |
| Language: | English |
| Published: |
Hibiscus Publisher
2014
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| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/10668/1/Over%20expression%20of%20Recombinant%20CP3%20%28abstract%29.pdf http://ir.unimas.my/id/eprint/10668/ http://journal.hibiscuspublisher.com/index.php/BSTR/article/view/159 |
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| Institution: | Universiti Malaysia Sarawak |
| Language: | English |
| Summary: | Currently, serological based diagnosis of rice tungro disease (RTD) is being limited by the
availability of antisera. This study aims to produce recombinant protein from the coat protein
(CP)3 of rice tungro spherical virus (RTSV) as a potential antigen in producing high titre
antibodies for use in detection of RTD. Amplified RTSV CP3 gene of approximately 900 bp
was cloned into pETSUMO vector. The recombinant plasmid was transformed into cloning host
Mach1™-T1® E.coli cells and screened for recombinant gene in the correct orientation. Plasmid
DNA from positive transformants was isolated and transformed into expression host BL21
(DE3) E.coli cells. One positive clone was selected for large scale expression where the
expressed protein was purified and analysed by SDS-PAGE and Western blot. This
recombinant RTSV CP3 protein, 46 kDa in molecular mass with 6x His-tag and SUMO protein
fused to its N-terminal, was found to be antigenic when it reacted with our in-house generated
polyclonal antibodies against tungro viruses. |
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