Characterization of theHelicoverpa assulta nucleopolyhedrovirus genome and sequence analysis of the polyhedrin gene region |
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Authors: | Soo-Dong Woo Jae Young Choi Yeon Ho Je Byung Rae Jin |
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Institution: | (1) Department of Plant Medicine, College of Agriculture, Life and Environment Sciences, Chungbuk National University, 361-763 Cheongju, Korea;(2) School of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, 151-742 Seoul, Korea;(3) College of Natural Resources and Life Science, Dong-A University, 604-714 Busan, Korea |
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Abstract: | A local strain ofHelicoverpa assulta nucleopolyhedrovirus (HasNPV) was isolated from infectedH. 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 kbEcoRI, 15 kbNcoI, 20 kbXhoI, 17 kbBgl II and 3 kbClaI fragments, respectively. The 3 kbClaI 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 fromAutographa californica NPV but were most closely related toHelicoverpa andHeliothis species NPVs with over 99% sequence identity. |
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Keywords: | Baculovirus genome HasNPV Helicoverpa assulta polyhedrin gene |
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