BEGIN:VCALENDAR
VERSION:2.0
PRODID:-//wp-events-plugin.com//6.6.4.4//EN
TZID:Asia/Jerusalem
X-WR-TIMEZONE:Asia/Jerusalem
BEGIN:VEVENT
UID:964@biotech.technion.ac.il
DTSTART;TZID=Asia/Jerusalem:20201012T170000
DTEND;TZID=Asia/Jerusalem:20220202T164952
DTSTAMP:20220512T124720Z
URL:https://biotech.technion.ac.il/events/analysis-of-the-alternative-sigm
 a-factor-system-%cf%83i5-rsgi5-in-clostridium-thermocellum-2/
SUMMARY:Analysis of the alternative sigma factor system\, σI5/RsgI5\, in C
 lostridium thermocellum
DESCRIPTION:Clostridium (Ruminiclostridium) thermocellum is a Gram-positive
 \, thermophilic\, anaerobic bacterium that is capable of efficiently hydro
 lyzing crystalline cellulose via an extracellular multienzyme complex\, te
 rmed the cellulosome. C. thermocellum regulates its cellulosomal genes in 
 response to extracellular biomass composition via a unique σI/RsgI biomas
 s sensing system. Unlike the previously characterized sigI-rsgI operons th
 at contain two genes\, the uncharacterized sigI5-rsgI5 operon is composed 
 of three genes including a gene upstream to sigI5\, for a putative arabino
 furanosidase from glycoside hydrolase family 43 (GH43). Transcription anal
 ysis using RT-PCR confirmed that gh43\, sigI5 and rsgI5 are co-transcribed
 . The apparent transcription start site of the gh43-rsgI5 operon was ident
 ified and conserved sequence in the promoter region was observed. The pred
 icted σI5 promoter sequence was identified in three arabino-xylan utiliza
 tion genes\, suggesting that their expression is controlled by σI5. Inter
 estingly\, the predicted σI5 promoter sequence was also identified in the
  upstream region of rsgI5\, suggesting a different type of gene organizati
 on and regulation of the gh43-rsgI5 operon compared to the other sigI-rsgI
  operons. Due to the limited genetic tools available for C. thermocellum\,
  a heterologous B. subtilis 168 strain host system was applied to verify t
 he functionality of the predicted promoters regulated by σI5. We were abl
 e to integrate C. thermocellum sigI5 gene and the putative σI5 regulated 
 promoters (fused to a reporter gene) into B. subtilis at different chromos
 omal sites\, and found that using multimers plasmids contributed to the su
 ccess of the integration events. Unfortunately\, we could not observe expr
 ession form the tested promoters. Real-time reverse transcription (RT) PCR
  revealed the lack of the sigI5 transcript in B. subtilis. We attempted to
  use a cell-free system\, but failed to obtain a soluble of the σI5 prote
 in in E. coli.\n\n \n\n 
END:VEVENT
BEGIN:VTIMEZONE
TZID:Asia/Jerusalem
X-LIC-LOCATION:Asia/Jerusalem
BEGIN:DAYLIGHT
DTSTART:20200327T030000
TZOFFSETFROM:+0200
TZOFFSETTO:+0300
TZNAME:IDT
END:DAYLIGHT
BEGIN:STANDARD
DTSTART:20201025T010000
TZOFFSETFROM:+0300
TZOFFSETTO:+0200
TZNAME:IST
END:STANDARD
BEGIN:DAYLIGHT
DTSTART:20210326T030000
TZOFFSETFROM:+0200
TZOFFSETTO:+0300
TZNAME:IDT
END:DAYLIGHT
BEGIN:STANDARD
DTSTART:20211031T010000
TZOFFSETFROM:+0300
TZOFFSETTO:+0200
TZNAME:IST
END:STANDARD
END:VTIMEZONE
END:VCALENDAR