cDNA cloning and sequence analysis of NIb gene of soybean mosaic virus.

Abstract

cDNA of soybean mosaic virus (Beijing isolate, SMV-BJ) has been synthesized, using viral genomic RNA as a template and random hexanucleotides as primers. Based on the sequences of SMV-BJ coat protein (CP) gene as well as SMV- and WMV-II-related regions, oligonucleotides were made as primers for polymerase chain reaction (PCR). NIb gene of SMV-BJ was amplified by PCR, and cloned into pBluescript SK. The complete sequence was determined. The comparison of NIb genes between SMV-BJ and WMV-II (USA) shows that similarities for nucleotide sequence reach 80.3%; and the deduced amino acid sequence, 91.3%. In consideration of the high identities in between the CP gene and the 3'-non-coding region between them, WMV-II might be considered as a watermelon strain of SMV. Besides, some unexpected sequences were found in the 3'-region of 2 NIb gene clones. Following modification and splicing, a binary vector of NIb gene has been constructed for its expression in higher plant for the purpose of studying the possible replicase-mediated resistance in polyviruses.