This is a generator of fcsR, that is used to regulate the level of c-di-GMP.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 7
Illegal NheI site found at 30
- 21Illegal XhoI site found at 766
- 23COMPATIBLE WITH RFC
- 25Illegal NgoMIV site found at 124
Illegal NgoMIV site found at 869
- 1000Illegal BsaI site found at 763
Illegal BsaI.rc site found at 511
Illegal SapI.rc site found at 876
This is a generator of fcsR (BBa_K3740022), that is used to regulate the level of c-di-GMP.
The coding 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.
As shown in Figure 2, the generator was identified successfully by PCR amplification.
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.