The p10 gene of Bombyx mori nucleopolyhedrosis virus encodes a 7.5-kDa protein and is hypertranscribed from a TAAG motif

Palhan, Vikas B and Gopinathan, Karumathil P (2000) The p10 gene of Bombyx mori nucleopolyhedrosis virus encodes a 7.5-kDa protein and is hypertranscribed from a TAAG motif. In: Journal of Genetics, 79 (2). pp. 33-40.

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Abstract

In baculovirus-based high-level expression of cloned foreign genes, the viral very late gene promoters of polyhedrin (polh)and p10 are extensively exploited. Here we report the cloning and characterization of the p10 gene from a local isolate of Bombyx mori nucleopolyhedrosis virus (BmNPV). The gene harbours a 213-bp open reading frame encoding a protein of 70 amino acids with a predicted molecular mass of 7.5 kDa. The BmNPV p10 showed deletion of a single A at á210 nucleotide compared to the prototype baculovirus, Autographa californica multinucleocapsid nucleopolyhedrosis virus (AcMNPV), p10 gene, resulting in a translational frameshift to generate a termination codon and consequently a truncated polypeptide instead of the 10-kDa protein. This protein P7.5 from BmNPV has a putative leucine zipper dimerization motif towards the N-terminal end and the central nuclear disintegration domain but the carboxy-terminal domain implicated in protein association for Æbrillar structure formation is absent.Phylogenetic analysis revealed that p10 is highly conserved among baculoviruses and the BmNPV strains are more closely related to AcMNPV than other baculoviruses.Thetranscription of p10 is regulated in a temporal manner, reaching maximal levels by 72 h post-infection. RNAase protection and primer extension analysis mapped the transcription start sites at ¡70 and ¡71 nt with respect to the ATG, within the conserved baculovirus late gene motif TAAG.The upstream region showed complete homology to the strong promoter of the AcMNPV p10, suggesting that this promoter from BmNPV could also be exploited for high-level expression of cloned foreign genes in silkworm cells or larvae.

Item Type:	Journal Article
Publication:	Journal of Genetics
Publisher:	Indian Academy of Sciences
Additional Information:	Copyright for this article belongs to Indian Academy of Sciences.
Keywords:	baculovirus;Bombyx mori;BmNPV;late gene expression;p10 gene;transcriptional regulation
Department/Centre:	Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited:	09 Sep 2004
Last Modified:	19 Sep 2010 04:16
URI:	http://eprints.iisc.ac.in/id/eprint/1953

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