RNA polymerase sigma-related proteins in Escherichia coli: detection by antibodies against a synthetic peptide |
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Authors: | Nobuyuki Fujita Akira Ishihama Yoji Nagasawa Susumu Ueda |
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Affiliation: | (1) Department of Molecular Genetics, National Institute of Genetics, 411 Mishima, Shizuoka, Japan;(2) Section of Immunochemistry and Biotechnology, Nippon Institute for Biological Science, 198 Ohme, Tokyo, Japan |
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Abstract: | Summary Antibodies were raised against a synthetic tetradecameric peptide with an amino acid sequence, DLIQEGNIGLMKAV, which corresponds to the most highly conserved region of bacterial RNA polymerase factors. In a Western-blot analysis of total Escherichia coli proteins, the antiserum reacted specifically with at least three proteins with apparent molecular weights of 75 kDa, 27 kDa and 23 kDa, in addition to the known factors (70 and 32). The majorities of 70 and 32 were recovered as associated forms with the RNA polymerase on glycerol gradient centrifugation, while the other cross-reacting proteins were not. Unambiguous evidence was obtained which indicated that the intracellular level of 32 increased rapidly upon heatshock, at least in the strain containing high copy numbers of the rpoH gene. |
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Keywords: | Escherichia coli RNA polymerase Sigma factor Heat-shock Synthetic peptide |
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