Part:BBa_K4765121

ribozyme connected: galU + pgmA + mScarlet

Introduction

This composite part is designed for the production of exopolysaccharide (EPS), and detection of EPS’s adhesion ability. It includes E. coli galU on the upstream site and E. coli pgmA on the downstream site, two essential enzymes in EPS biosynthesis pathway . These two enzymes were tested by iGEM20_XJTU-China in K3331001(galU)and K3331002(pgmA). The following mScarlet is utilized to detect E. coli 's adhesion ability. Additionally, during the validation of EPS, we observed that EPS expressing bacteria became "heavier" and "stickier", thereby promoting cluster formation.

Usage and Biology

We use this part to improve EPS production of E. coli and assessment of EPS’s adhesion ability.

Characterization

Agarose gel electrophoresis

Figure 1. Agarose gel electrophoresis of PCR products, amplified from bacterial colonies/cultures. From left lane(1) to right lane(3) indicate the successful construction of galU, galU + pgmA, and galU + pgmA + mScarlet.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NotI site found at 3285
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]