PMID:9893021

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Citation

Trigwell, S and Glass, RE (1998) Function in vivo of separate segments of the beta subunit of Escherichia coli RNA polymerase. Genes Cells 3:635-47

Abstract

Transcription of genetic material is catalysed by the enzyme DNA-dependent, RNA polymerase. The multimeric RNA polymerases consist of between 4 and 16 different subunits, of which the two largest, termed beta and beta', are conserved throughout nature. The beta subunit has been implicated in all of the stages of transcription that are catalysed by the complete enzyme. Several lines of evidence have suggested that the function of the beta subunit is not dependent upon the contiguity of the sequence blocks. In this report, a complementary immunological and genetic approach was adopted in order to investigate the individual regions of the beta subunit of RNA polymerase. To this end, the beta structural gene rpoB was separated into four near-equal, non-overlapping segments (as well as 'half' genes) on the basis of 'split' genes in nature, known functional organization and sequence conservation. These segments were used to prepare sequence-specific antibodies against the four individual regions, as well as being expressed in vivo from a tight, lac-controlled high-copy number vector.

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Keywords

Animals; DNA-Directed RNA Polymerases/chemistry; DNA-Directed RNA Polymerases/genetics; DNA-Directed RNA Polymerases/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Genes, Bacterial; Genetic Complementation Test; Peptide Elongation Factor Tu/metabolism; Precipitin Tests; Rabbits

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