Antibody Directed against Bacillus subtiLis CT Factor Purified by Sodium Dodecyl Sulfate Slab Gel Electrophoresis EFFECT ON TRANSCRIPTION BY RNA POLYMERASE IN CRUDE EXTRACTS OF VEGETATIVE AND SPORULATING CELLS*
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چکیده
Antibody directed against c factor from vegetative Bacillus subtilis was prepared by immunizing a rabbit with denatured cr polypeptide isolated by electrophoresis of partially purified DNA-dependent RNA polymerase on a sodium dodecyl sulfate-polyacrylamide slab gel. Antiserum to (r reacted specifically with native c polypeptide but not with core RNA polymerase as judged by complement fixation and by an immunodiffusion assay. Anti-c antibody also inhibited the ability of a to stimulate transcription of phage be DNA but failed to inhibit transcription of poly(dA-dT) by core enzyme. Specific antibody was also raised against a mixture of the p and p’ subunits of RNA polymerase purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The effect of the anti-o y-globulin on the transcription of phage @e DNA by RNA polymerase in crude extracts of vegetative and sporulating cells was examined. Anti-u antibody markedly inhibited the transcription of phage DNA by RNA polymerase partially purified from vegetative bacteria by ammonium sulfate fractionation but had little effect on transcription of the phage DNA template by enzyme from sporulating cells. Addition of purified d to a vegetative extract that had been depleted of u by treatment with the anti-a antibody restored active transcription of phage DNA. However, addition of purified v to an antibody-treated extract of sporulating cells had little effect on @e RNA synthesis. These findings suggest that sporulating cells contain a component that interferes with the activity of the CJ subunit of RNA polymerase.
منابع مشابه
Isolation of a new RNA polymerase-binding protein from sporulating Bacillus subtilis.
RNA polymerase was precipitated from extracts of radioactively labeled vegetative and sporulating Bacillus subtilis with antiserum prepared against vegetative core polymerase. The precipitates were solubilized and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Antiserum added to an extract of vegetative B. subtilis precipitated only the known subunits of core RNA polymer...
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