Construction of Plant Virus Vector of Tobacco Necrosis Virus

An almost complete sequence of the RNA genome of tobacco necrosis virus(TNV)strain A has been determined.Tobacco necrosis virus A is ssRNA virus.It contains six open reading frames.SphI was constructed between ORFⅣ and ORFⅤ(coat protein)of TNV-A gene that has been cloned into pMD18-T vector by overlapping extension of PCR.The pE1802-TNV of plant expression vector was constructed with the mutated TNV-A gene inserted into SalI and SmaI[WTBZ] of pE1802.