Thermococcales Transcription Initiation Response to Stress Conditions
Presentation Type
Poster
Start Date
5-4-2022 11:00 AM
End Date
5-4-2022 1:00 PM
Subjects
Archaea, Transcription Initiation, Thermophiles, Thermococcales
Advisor
Dr. Mike Bartlett
Student Level
Masters
Abstract
Pyrococcus furiosus is an anaerobic thermophilic Archaean within Thermococcales living in shallow sea hydrothermal vents. This species has two transcription initiation factors: Transcription Factor B (TFB) and Transcription Factor B 2 (TFB2). TFB is essential and is homologous to Eukaryotic TF2B. TFB2 is missing the DNA binding B-factor domain resulting in poor DNA interaction. Unknown conditions cause TFB2 to be produced. Pyrococcus woesei and P. furiosus are, controversially, two species but only P. furiosus contains TFB2. P. woesei performs significantly worse under identical oxidative stress conditions compared to P. furiosus, (Strand, 2010). NAP is a sequenced but unevaluated protein adjacent to TFB2 found in P. furiosus and Thermococcus PK. TFB2 has a weak connection to heat stock response, (Shockley 2003). A P. furiosus deletion of TFB2 will be made using a popout sequence during Gibson assembly allowing for reinsertion of the deleted sequence. This verifies identical behavior to the wild type and that genetic scars don’t have ulterior interference. Over expressions of NAP will be produced for a transcription assay. Survivability in P. furiosus (TFB2ΔTFB2, ΔTFB2, and wild type), Thermococcus PK, and P. woesei will be measured under variable oxidative and heat stress to evaluate their combined impact.
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https://archives.pdx.edu/ds/psu/37499
Thermococcales Transcription Initiation Response to Stress Conditions
Pyrococcus furiosus is an anaerobic thermophilic Archaean within Thermococcales living in shallow sea hydrothermal vents. This species has two transcription initiation factors: Transcription Factor B (TFB) and Transcription Factor B 2 (TFB2). TFB is essential and is homologous to Eukaryotic TF2B. TFB2 is missing the DNA binding B-factor domain resulting in poor DNA interaction. Unknown conditions cause TFB2 to be produced. Pyrococcus woesei and P. furiosus are, controversially, two species but only P. furiosus contains TFB2. P. woesei performs significantly worse under identical oxidative stress conditions compared to P. furiosus, (Strand, 2010). NAP is a sequenced but unevaluated protein adjacent to TFB2 found in P. furiosus and Thermococcus PK. TFB2 has a weak connection to heat stock response, (Shockley 2003). A P. furiosus deletion of TFB2 will be made using a popout sequence during Gibson assembly allowing for reinsertion of the deleted sequence. This verifies identical behavior to the wild type and that genetic scars don’t have ulterior interference. Over expressions of NAP will be produced for a transcription assay. Survivability in P. furiosus (TFB2ΔTFB2, ΔTFB2, and wild type), Thermococcus PK, and P. woesei will be measured under variable oxidative and heat stress to evaluate their combined impact.