Turnip mosaic virus coat protein gene: Cloning and construction of the plant vector

Kong, L.-Jie; Fang, R.-Xiang; Chen, Z.-Hua; Mang, K.-Qiang

Science in China Series B Chemistry Life Sciences and Earth Sciences 35(12): 1444-1452


Accession: 009681940

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Complementary DNAs to turnip mosaic virus (a ridish Raphanus stativus isolate) genomic RNA were synthesized using oligo(dT) as primer and cloned into the vector lambda-ZAP II. After hybridization with a single-stranded cDNA probe and the sequencing of inserted DNA, positive clones with poly-A tails were obtained. One clone containing 1429-base pair insert was sequenced. The coat protein gene was identified based on the molecular weight of the TuMV coat protein and the consensus sequences of the polyprotein processing sites of potyviruses. The 5' end of the coat protein gene was modified by PCR to introduce an initiation codon, ATG, and two restriction enzyme sites. The gene was then manipulated into a binary vector pBIN437 which was derived from pBI121, and the plant expression vector is being used to transform Brassica napus.