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Molecular cloning, sequence analysis and expression of genome segment 7 (S7) of Antheraea mylitta cypovirus (AmCPV) that encodes a viral structural protein

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Abstract

The Genome segment 7 (S7) of the 11 double stranded RNA genomes from Antheraea mylitta cypovirus (AmCPV) was converted to cDNA, cloned and sequenced. The nucleotide sequence showed that segment 7 consisted of 1789 nucleotides with an ORF of 530 amino acids and could encode a protein of approximately 61 kDa, termed P61. The 5′ terminal sequence, AGTAAT and the 3′ terminal sequence, AGAGC of the plus strand was found to be the same as genome segment 10 of AmCPV encoding polyhedrin. No sequence similarity was found by searching nucleic acid and protein sequence databases using BLAST. The secondary structure prediction showed the presence of 17 α-helices, 18 extended β-sheets along the entire length of P61. The ORF of segment 7 was expressed in E. coli as His-tagged fusion protein, purified through Ni-NTA chromatography, and polyclonal antibody was raised in rabbit indicating that P61 is immunogenic. Immunoblot analysis using this antibody on viral infected cells as well as purified polyhedra showed that P61 is a viral structural protein. Motif scan search showed some similarity of P61 with Inosine monophosphate dehydrogenase (IMPDH) cystathionine-β-synthase (CBS) domain at the C-terminus and it was hypothesized that by binding to single stranded viral RNA through its CBS domain P61 may help in virus replication or transcription.

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Acknowledgments

The authors thank the Director(s) of Central Tasar Research and Tranining Institute, Ranchi and Jhargram for providing the virus infected A. mylitta larvae. Venkata Ramana Murthy Chavali is the recipient of fellowship from Indian Institute of Technology, Kharagpur and Council of Scientific and Industrial Research, New Delhi.

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  1. Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur, 721302, India

    Venkata Ramana Murthy Chavali & Ananta K. Ghosh

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  1. Venkata Ramana Murthy Chavali
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  2. Ananta K. Ghosh
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Correspondence to Ananta K. Ghosh.

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Chavali, V.R.M., Ghosh, A.K. Molecular cloning, sequence analysis and expression of genome segment 7 (S7) of Antheraea mylitta cypovirus (AmCPV) that encodes a viral structural protein. Virus Genes 35, 433–441 (2007). https://doi.org/10.1007/s11262-006-0070-z

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