IIsolation of GAG-CA Subunit of Jembrana Virus from the Viral Genome by RT-PCR and its cloning in pCR2.1-topolasmid using topoisomerase-based system
Jembrana virus is agent of acute Jembrana virus disease (JVD) in Bos javanicus. The genome of Jembrana virus is composed of a single-stranded RNA. We described in this paper the isolation of gag-ca subunit gene from the viral genome by a single-step RT-PCR reaction. The isolated gene was inserted in...
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| Format: | Article NonPeerReviewed |
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[Yogyakarta] : Universitas Gadjah Mada
2002
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| Online Access: | https://repository.ugm.ac.id/18409/ http://i-lib.ugm.ac.id/jurnal/download.php?dataId=1195 |
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| Institution: | Universitas Gadjah Mada |
| Summary: | Jembrana virus is agent of acute Jembrana virus disease (JVD) in Bos javanicus. The genome of Jembrana virus is composed of a single-stranded RNA. We described in this paper the isolation of gag-ca subunit gene from the viral genome by a single-step RT-PCR reaction. The isolated gene was inserted in pCR2.1-TOPO plasmid by TOPO TA cloning kit, based on topoisomerase system. It allowed the cloning procedure using the RT-PCR products without prior purification or restriction enzyme digestion. A high number of positive recombinant bacterial colonies were obtained which were further analysis by BamHI-EcoRI digestion for a definite characterization of positive clones. The cloning procedure we used is highly efficient enabling rapid and easy gene isolation.
Keywords: Jembrana virus disease- gag ca gene - RT-PCR-Topoisomerase cloning |
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