[Studies on selectivity of recognizing factor for rmf promoter]

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.