Characterization of the Helicoverpa assulta nucleopolyhedrovirus genome and sequence analysis of the polyhedrin gene region.
A local strain of Helicoverpa assulta nucleopolyhedrovirus (HasNPV) was isolated from infected H.assulta larvae in Korea. Restriction endonuclease fragment analysis, using 4 restriction enzymes, estimated that the total genome size of HasNPV is about 138 kb. A degenerate polymerase chain reaction (PCR) primer set for the polyhedrin gene successfully amplified the partial polyhedrin gene of HasNPV. The sequencing results showed that the about 430 bp PCR product was a fragment of the corresponding polyhedrin gene. Using HasNPV partial predicted polyhedrin to probe the Southern blots, we identified the location of the polyhedrin gene within the 6 kb Eco RI, 15 kb Nco I, 20 kb Xho I, 17 kb Bgl II and 3 kb Cla I fragments, respectively. The 3 kb Cla I fragment was cloned and the nucleotide sequences of the polyhedrin coding region and its flaking regions were determined. Nucleotide sequence analysis indicated the presence of an open reading frame of 735 nucleotides which could encode 245 amino acids with a predicted molecular mass of 29 kDa. The nucleotide sequences within the coding region of HasNPV polyhedrin shared 73.7% identity with the polyhedrin gene from Autographa californica NPV but were most closely related to Helicoverpa and Heliothis species NPVs with over 99% sequence identity.
Woo
Soo-Dong
SD
Department of Plant Medicine, College of Agriculture, Life and Environment Sciences, Chungbuk National University, Cheongju 361-763, Korea. sdwoo@cbnu.ac.kr
Choi
Jae Young
JY
Je
Yeon Ho
YH
Jin
Byung Rae
BR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D016679:Genome, Viral; D007814:Larva / Q000821:virology; D007915:Lepidoptera / Q000821:virology*; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000302:isolation & purification; D000077183:Occlusion Body Matrix Proteins; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000145:classification / Q000235:genetics*; D015678:Viral Structural Proteins
D000077183:Occlusion Body Matrix Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus