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Part:BBa_K3740021
rocR, PA3947, cyclic di-GMP phosphodiesterase
Description
Degradation of c-di-GMP in Gluconacetobacter hansenii ATCC 53582.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 258
Illegal BamHI site found at 1171
Illegal XhoI site found at 94 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 895
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 802
2021 SZPT-China
Biology
The gene rocR is originated from the genome of Pseudomonas aeruginosa (PAO1). And RocR expressed by rocR is the phosphodiesterase (PDE) of c-di-GMP.
Usage
The encoding sequences for RocR were inserted into the expression vector with BBa_K880005 (BBa_J23100 & BBa_B0034) to obtain J23100-B0034-rocR-rrnB T1 (BBa_K3740065). 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.
Characterization
1. Identification
<p>As shown in Figure 2, c-di-GMP phosphodiesterase-encoded genes BBa_K3740065 was identified successfully by PCR amplification.![](/wiki/images/4/41/Szpt16.png)
2. Characterization
As shown in Figure 3, BC production in J23100-B0034-rocR-rrnB T1-pSEVA331- G. hansenii ATCC 53582 and the control group pSEVA331-G. hansenii ATCC 53582 was not significantly different, indicating that RocR was not capable of hydrolyzing c-di-GMP in G. hansenii ATCC 53582.
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