Part:BBa_K318502

lacI pL + RBS + RcsB + RBS + RcsA + TT

Our goals is to have each cell produce enough colonic acid to protect itself from the harsh acidic conditions of the stomach. We placed RcsA, RcsB, and a combination of the two under a IPTG inducible promoter to test both quantity of colonic acid produced and cell survivability. You can see our data by downloading the associated .zip file for any of these three devices:BBa_K318500(RcsA), BBa_K318501(RcsB), BBa_K318502(RcsA+RcsB)

Usage and Biology

RcsA and RcsB are transcription factors that are positive regulator of capsular polysaccharide synthesis. The two transcription factors form a heterodimer that is know to activate around 19 genes related to colonic acid synthesis. RcsB is also know to form a homodimmer and positivly regulate cell division. RcsA and RcsB belong to the multicomponent RcsF/RcsC

See more on http://biocyc.org/ECOLI/NEW-IMAGE?type=GENE&object=EG10820 Ecocyc-RcsA & http://biocyc.org/ECOLI/NEW-IMAGE?type=GENE&object=EG10821 Ecocyc-RcsB

Characterization

Zip File Download media:BBa_K318502_pSB1C3.zip

Figure 1: L-Fucose content of plasmids K318500, K318501, and K200021 (control) in backbone pSB1AK3 transformed in MG1655. Colonic Acid is a polysaccharide containing a repeat unit with D-glucose, L-fucose, D-galactose, and D-glucuronate. Colonic acid can be estimated by measuring L-fucose content.

Figure 2: Survivability of plasmids K318500, K318501, K318502, and K200021 (control) in backbone pSB1AK3 transformed in MG1655. We saw around a 10x increase in survivability with RcsB being over produced at a pH=4.

For more detail on testing, please see our results page on our Team Wiki for protocols, background information, and conclusions.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]