Difference between revisions of "Part:BBa K3740021"
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<h4>1. Identification</h4> | <h4>1. Identification</h4> | ||
<p>As shown in Figure 2, c-di-GMP phosphodiesterase-encoded genes <partinfo>BBa_K3740065</partinfo> was identified successfully by PCR amplification.</p> | <p>As shown in Figure 2, c-di-GMP phosphodiesterase-encoded genes <partinfo>BBa_K3740065</partinfo> was identified successfully by PCR amplification.</p> | ||
+ | [[File:szpt16.png|200px|thumb|center|Figure 2. Agarose gel electrophoresis image of J23100-B0034-rocR-rrnB T1 (<partinfo>BBa_K3740065</partinfo>). BBa_K3740065, 1294bp]] | ||
+ | <h4>2. Characterization</h4> | ||
+ | <p>As shown in Figure 3, BC production in J23100-B0034-rocR-rrnB T1-pSEVA331- <i>G. hansenii</i> ATCC 53582 and the control group pSEVA331-<i>G. hansenii</i> ATCC 53582 was not significantly different, <b>indicating that RocR was not capable of hydrolyzing c-di-GMP in <i>G. hansenii</i> ATCC 53582.</b></p> | ||
+ | [[File:szpt17.png|200px|thumb|center|Figure 3. BC yield by J23100-B0034-rocR-rrnB T1-pSEVA331-<i>G. hansenii</i> ATCC 53582 and the vehicle control pSEVA331-<i>G. hansenii</i> ATCC 53582.]] |
Revision as of 14:18, 18 October 2021
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.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.