Cloning and sequencing of the cDNA encoding for pokeweed anti-viral protein (PAP) and construction of its plant expression vector

Zhang HaiYan; Liu GuiZhen; Chen ZhengHua

Acta Botanica Sinica 41(2): 226-228

1999


ISSN/ISBN: 0577-7496
Accession: 003384801

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Abstract
Total RNA was extracted from the leaves of pokeweed (Phytolacca americana) plants which had been subcultured for 8 weeks. The first strand of cDNA was synthesized from the total RNA template with oligo(dT)15 primer using MMLV reverse transcriptase. A 0.96 kb cDNA fragment was obtained after 30 PCR amplification cycles with two specific primers. The cDNA fragment was sequenced from two directions after being cloned into pGEM-T Easy Vector. The results showed that the cDNA clone to a previously published pokeweed anti-viral protein (PAP) cDNA clone. The plant expression vector with the PAP cDNA was constructed, and the work of transferring the PAP cDNA into tobacco and rape to obtain transgenic plants resistant to a broadspectrum of viruses is in progress.