商陆抗病毒蛋白cDNA的克隆、测序及其植物表达载体的构建

The total RNA was extraeted from the leaves of pokeweed (phytolacca americana L.) plants whichhad been subcultured for eight weeks. The first stran of eDNA was synthesized from the total RNA templatewith oligo(dT) 15 primer using MMLV reverse transeriptase.A 0. 96 kb cDNA fragment was obtained after 30PCR amplification cycles with two speeifie primers. The cDNA fragment was sequenced from from directions after being cloned into pGEM-T Easy Veetor.The result showed that the cDNA clone had the entire coding region with the same sequence as the previously published pokeweed anti-viral protein (PAP)cDNA clone. Theplant expression vector with the PAP cDNA was eonstrueted,and the work of transfekrring the PAP cDNA intotobacco and rape obtain transgenie plants resistant to a broadspectrum of viruses in in progress.