Designed recombinant transcription factor with antibody-variable regions.
Many recombinant transcription factors have been invented, but we cannot select a substance used as an inducer. In this study, we have created a novel expression control system in which we can select a substance as an inducer toward which a monoclonal antibody (mAb) is prepared. The variable region fragments (Fvs) of the heavy and light chains (V(H) and V(L)) of the bisphenol A (BPA)-specific mAb BBA-2187 were each fused to the DNA-binding domain (DBD) of LexA and the transactivation domain (AD) of VP16. The association between the two recombinant proteins in the presence of BPA constituted a functional transcription factor. The recombinant proteins in which the DBD was fused to the N-terminal side of the Fv and in which the nuclear localization signal (NLS) was fused to the N-termini of the construct including the AD highly induced beta-galactosidase (lacZ) expression in recombinant yeast cells grown with BPA. When the Fvs of the polychlorinated biphenyl (PCB)-specific mAb 4444 were used, DBD-NLS-V(H) and NLS-AD-V(L) showed significantly increased lacZ activity in response to a PCB derivative. The Fv transcription factor may be useful in many fields such as gene therapeutics.
Gion
Keiko
K
Research Center for Environmental Genomics, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, Hyogo 657-8501, Japan.
Sakurai
Yuki
Y
Watari
Akito
A
Inui
Hideyuki
H
D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000737:chemistry / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology; D001559:Benzhydryl Compounds; D004269:DNA, Bacterial / Q000737:chemistry / Q000276:immunology; D005786:Gene Expression Regulation; D017405:Herpes Simplex Virus Protein Vmw65 / Q000737:chemistry / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000276:immunology*; D007763:Lac Operon / Q000235:genetics; D010636:Phenols / Q000737:chemistry / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000737:chemistry / Q000276:immunology; D014157:Transcription Factors / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D000099160:LexA Repressor Protein; D000098826:Bisphenol A Compounds
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D001426:Bacterial Proteins; D001559:Benzhydryl Compounds; D004269:DNA, Bacterial; D017405:Herpes Simplex Virus Protein Vmw65; D007135:Immunoglobulin Variable Region; D010636:Phenols; D011994:Recombinant Proteins; D012697:Serine Endopeptidases; D014157:Transcription Factors; D000099160:LexA Repressor Protein; C006780:bisphenol A; D000098826:Bisphenol A Compounds