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Related Experiment Videos

[Studies on selectivity of recognizing factor for rmf promoter]

Q Ding1, I Akira

  • 1Wuhan Institute of Virology, Academia Sinica.

Wei Sheng Wu Xue Bao = Acta Microbiologica Sinica
|February 1, 1997
PubMed
Summary

This study shows that the rmf gene promoter is transcribed by E. coli RNA polymerase holoenzyme (E sigma) with sigma 70, but not sigma 38. Optimal transcription conditions were identified.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The rmf gene plays a role in bacterial physiology.
  • Bacterial transcription initiation is mediated by RNA polymerase holoenzyme.

Purpose of the Study:

  • To investigate the transcription of the rmf gene promoter in vitro.
  • To determine which sigma factors are involved in rmf gene transcription.
  • To optimize in vitro transcription conditions.

Main Methods:

  • In vitro transcription using E. coli RNA polymerase holoenzyme (E sigma) reconstituted with core enzyme and sigma 70 or sigma 38.
  • Identification of transcription initiation sites using restriction endonucleases.

Main Results:

  • The rmf promoter was recognized and transcribed by E. coli RNA polymerase holoenzyme containing sigma 70 (E sigma 70).
  • The rmf promoter was not recognized by E. coli RNA polymerase holoenzyme containing sigma 38 (E sigma 38).
  • Optimal in vitro transcription occurred at 37°C and 50 mmol/L NaCl.

Conclusions:

  • Sigma factor 70 is essential for the in vitro transcription of the rmf gene promoter.
  • Sigma factor 38 does not participate in rmf gene transcription.
  • Specific temperature and salt concentrations are crucial for efficient in vitro transcription.

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