Part:BBa_K3740022

fcsR, PA2133, cyclic di-GMP phosphodiesterase

Description

Degradation of c-di-GMP in Gluconacetobacter hansenii ATCC 53582.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 703
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 61
  Illegal NgoMIV site found at 806
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 700
  Illegal BsaI.rc site found at 448
  Illegal SapI.rc site found at 813
  

2021 SZPT-China

Biology

fcsR is originated from the genome of Pseudomonas aeruginosa (PAO1), the FcsR expressed by fcsR is the phosphodiesterase (PDE) of c-di-GMP.

Usage

The encoding sequences for FcsR were inserted into the expression vector with BBa_K880005 (BBa_J23100 & BBa_B0034) to obtain J23100-B0034-fcsR-rrnB T1 (BBa_K3740062). We introduced the constructed plasmid into E. coli DH5α to verify its successful construction, and then transferred it into G. hansenii ATCC 53582 to verify its function.

Figure 1. Gene circuit of fcsR.

Characterization

1. Identification

As shown in Figure 2, c-di-GMP phosphodiesterase-encoded genes BBa_K3740062 was identified successfully by PCR amplification.

Figure 2. Agarose gel electrophoresis image of J23100-B0034-fcsR-rrnB T1 (BBa_K3740062). BBa_K3740062, 975bp

2. Characterization

As shown in Figure 3, BC production in J23100-B0034-fcsR-rrnB T1-pSEVA331-G. hansenii ATCC 53582 was lower than the control group pSEVA331-G. hansenii ATCC 53582, indicating that FcsR was capable of hydrolyzing c-di-GMP in G. hansenii ATCC 53582.

Figure 3. BC yield by J23100-B0034-fcsR-rrnB T1-pSEVA331-G. hansenii ATCC 53582 and the vehicle control pSEVA331-G. hansenii ATCC 53582.