Molecular cloning and expression of key gene encoding hypothetical DNA polymerase from B. mori parvo-like virus

Molecular cloning and expression of key gene encoding hypothetical DNA polymerase from B. mori parvo-like virus

Blog Article

BmPLV-Z is the abbreviation for Bombyx mori parvo-like virus (China isolate).This is a novel virus with two single-stranded Wood Sign linear DNA molecules, viz., VD1 (6543 bp) and VD2 (6022 bp), which are encapsidated respectively into separate virions.

Analysis of the deduced amino acid sequence of VD1-ORF4 indicated the existence of a putative DNA-polymerase with exonuclease activity, possibly involved in the replication of BmPLV-Z.In the present study, a recombinant baculovirus was constructed to express the full length of the protein encoded by the VD1-ORF4 gene (3318 bp).In addition, a 2163-bp fragment amplified from the very same gene was cloned into prokaryotic expression vector pET-30a and expressed in E.

coli Rosetta 2 (DE3) pLysS.The expressed fusion protein was employed to immunize New Zealand white rabbits for the production of Cutlery Set an antiserum, afterwards used for examining the expression of the protein encoded by VD1-ORF4 gene in Sf-9 cells infected with recombinant baculovirus.Western blot analysis of extracts from thus cells infected revealed a specific band of about 120 kDa, thereby indicating that the full length protein encoded by the VD1-ORF4 gene had been successfully and stably expressed in Sf-9 cells.