cDNA cloning and sequence analysis of NIb gene of soybean mosaic virus.
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.
Liu
J
J
Institute of Microbiology, Chinese Academy of Sciences, Beijing.
Peng
X
X
Mang
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D002213:Capsid / Q000235:genetics; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D005814:Genes, Viral; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013025:Glycine max / Q000235:genetics / Q000821:virology*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics*
D017931:DNA Primers; D018076:DNA, Complementary; D014764:Viral Proteins; C061396:nuclear inclusion protein b, mosaic viruses; D012321:DNA-Directed RNA Polymerases