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Characterization of theHelicoverpa assulta nucleopolyhedrovirus genome and sequence analysis of the polyhedrin gene region

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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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Abbreviations

AcNPV:

Autographa californica nucleopolyhedrovirus

HasNPV:

Helicoverpa assulta nucleopolyhedrovirus

ORF:

open reading frame

PCR:

polymerase chain reaction

REN:

restriction endonuclease

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Authors and Affiliations

  1. Department of Plant Medicine, College of Agriculture, Life and Environment Sciences, Chungbuk National University, 361-763, Cheongju, Korea

    Soo-Dong Woo, Jae Young Choi, Yeon Ho Je & Byung Rae Jin

  2. School of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, 151-742, Seoul, Korea

    Jae Young Choi & Yeon Ho Je

  3. College of Natural Resources and Life Science, Dong-A University, 604-714, Busan, Korea

    Byung Rae Jin

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  1. Soo-Dong Woo
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  2. Jae Young Choi
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  3. Yeon Ho Je
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  4. Byung Rae Jin
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Correspondence to Soo-Dong Woo.

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Woo, SD., Choi, J.Y., Je, Y.H. et al. Characterization of theHelicoverpa assulta nucleopolyhedrovirus genome and sequence analysis of the polyhedrin gene region. J. Biosci. 31, 329–338 (2006). https://doi.org/10.1007/BF02704105

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