Temperature-sensitive Escherichia coli mutant producing a temperature-sensitive sigma subunit of DNA-dependent RNA polymerase. | Zendy

J. D. Harris | Zendy; Joseph S. Heilig | Zendy; I I Martinez | Zendy; Richard Calendar | Zendy; Leif A. Isaksson | Zendy

Open Access

Temperature-sensitive Escherichia coli mutant producing a temperature-sensitive sigma subunit of DNA-dependent RNA polymerase.

Author(s) -

J. D. Harris,

Joseph S. Heilig,

I I Martinez,

Richard Calendar,

Leif A. Isaksson

Publication year - 1978

Publication title -

proceedings of the national academy of sciences

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 5.011

H-Index - 771

eISSN - 1091-6490

pISSN - 0027-8424

DOI - 10.1073/pnas.75.12.6177

Subject(s) - microbiology and biotechnology , mutant , dnag , rna polymerase , biology , escherichia coli , specificity factor , sigma factor , transcription (linguistics) , protein subunit , gene , genetics , circular bacterial chromosome , linguistics , philosophy

A gene affecting the sigma subunit of DNA-dependent RNA polymerase is tightly linked to dnaG at 66 min on the Escherichia coli chromosome. In order to create an easily selectable marker in this region, we inserted transposon-10, which carries a gene determining resistance to tetracycline (tet) near 66 min, and the order tolC-dnaG-sigma-tet was determined. We used frequency of contransduction with tet as a criterion to screen a collection of spontaneous temperature-sensitive Escherichia coli mutants that might affect the sigma subunit. One such mutant was found to map at the sigma locus. The sigma subunit isolated from this mutant is unstable at 46 degrees C in vitro and has an altered electrophoretic mobility. The temperature sensitivity of RNA synthesis in this mutant indicates that most transcription in E. coli is sigma dependent.

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