Expression of human papillomavirus type 6b E2 gene product with DNA-binding activity in insect (Bombyx mori) cells using a baculovirus expression vector.
Human papillomavirus type 6b (HPV6b) has been shown to be a major etiologic agent of genital condylomas. The E2 gene, one of the early genes, has been shown to activate the enhancer element in trans in cells transformed with bovine papillomavirus type 1a (BPV1a) but the E2 gene product of any HPV has not been identified. The E2 gene of HPV6b was inserted into the polyhedrin gene of a Baculovirus, Bombyx mori nuclear polyhedrosis virus (BmNPV), 156 bp downstream from the translational start codon, and transferred into BmNPV by allelic replacement in a cotransfected Bombyx mori cell line, Bm-N. The predicted 46-kDa protein was produced by a recombinant virus in the infected Bm-N cells at a high level under the control of the polyhedrin promoter. We obtained the antibody against the putative E2-polyhedrin fusion protein by immunizing a rabbit with this protein. This protein reacted with the antibodies against polyhedrin and the fusion protein. This protein did specifically bind to the upstream regulatory region of the HPV6b and BPV1a genomes. This DNA binding activity was blocked by the antibody against this protein.
Sekine
H
H
Department of Microbiology, Chiba University, School of Medicine, Japan.
Fuse
A
A
Tada
A
A
Maeda
S
S
Simizu
B
B
D000818:Animals; D012831:Bombyx / Q000235:genetics; D002460:Cell Line; D004274:DNA, Recombinant; D004268:DNA-Binding Proteins / Q000235:genetics*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D007304:Insect Viruses / Q000235:genetics; D027383:Papillomaviridae / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*
D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins