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Synthesis and Cloning of cDNA Encoding an Antiviral Protein from the Leaves of Bougainvillea spectabilis Willd. (Nyctaginaceae)

Author(s): S. Rajesh | R. Balasaraswathi | Sabitha Doraisamy

Journal: World Journal of Agricultural Sciences
ISSN 1817-3047

Volume: 1;
Issue: 2;
Start page: 101;
Date: 2005;
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Keywords: Antiviral proteins BAP | cDNA synthesis | RT-PCR | Cloning | Homology search | CLUSTAL analysis

Total RNA was extracted from the mature leaves of Bougainvillea spectabilis Willd. and messenger RNA (mRNA) was separated out. Using mRNA as template, complementary DNA (cDNA) was synthesized and amplified by Reverse Transcription coupled PCR using gene specific primer. A product of 750 base pair plus was selected based on the size of Bougainvillea antiviral protein (BAP). After elution the product was purified and cloned into pGEM-T Easy vector and mobilized into E. coli strain, JM 109 and sequenced. The partial cDNA sequence of 893 base pairs contained three open reading frames. The sequence of a most possible ORF (297 bp) was translated and its homology with already reported sequences of antiviral proteins was analyzed.
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