Constrution of a near full - length cDNA library of wheat rosette stunt virus (WRSV)

A near full - length cDNA library of Wheat Rosette Stunt Virus (WRSV) was constructed.Purified viral RNA was polyadenylated by E. coli poly(A) polymerase. The poly(A) - tailed RNA was used as a template complementary to oligo dT(12 - 18) primer for cDNA synthesis.The cDNA was separately digested with restriction enzymes Pst I and EcoR I/Sma I. The pUC19 plasmid was digested with the same enzymes to poduce the both cohesive - ends, or one blunt and the other cohesive - ends respectively. Ten inserts with different sizes were obtained and the total size of them reached approximately 14 kb. One insert of near 6 kb length was identified as the insert containing the N protein gene and covering the Ns, M, G, genes as well as a part of L gene.The cDNA inserts with orientated - direction were sequenced and the sequence was compared to 5' and 3' ends of the genomic RNAs of two other plant rhabdoviruses, SYNV and LNYV using computer assisted searching method with the Software Sequence Analysis Version 3. 00.