Difference between revisions of "Part:BBa K3740019"
(→2021 SZPT-China) |
(→2021 SZPT-China) |
||
| Line 20: | Line 20: | ||
[[File:szpt18.png|400px|thumb|center|Figure 1. Engineering a potent photoactivated DGC]] | [[File:szpt18.png|400px|thumb|center|Figure 1. Engineering a potent photoactivated DGC]] | ||
<h3>Usage</h3> | <h3>Usage</h3> | ||
| − | The coding sequences for BphS and BphO were inserted into the expression vector with <partinfo>BBa_K880005</partinfo> (BBa_J23100 & BBa_B0034) to obtain J23100-B0034-bphS-bphO-rrnB T1 (BBa_K3740047). 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. | + | The coding sequences for BphS and BphO were inserted into the expression vector with <partinfo>BBa_K880005</partinfo> (<partinfo>BBa_J23100</partinfo> &<partinfo>BBa_B0034</partinfo>) to obtain J23100-B0034-bphS-pET RBS-bphO-rrnB T1 (<partinfo>BBa_K3740047</partinfo>). We introduced the constructed plasmid into <i>E. coli</i> DH5α to verify its successful construction, and then transferred it into<i> G. hansenii </i>ATCC 53582 to verify its function. |
| + | [[File:szpt11png|400px|thumb|center|Figure 2. Gene circuit of bphS]] | ||
Revision as of 13:26, 18 October 2021
bphS, photo-activated diguanylate cyclase
Description
Near-infrared light activated synthesis of c-di-GMP in Gluconacetobacter hansenii ATCC 53582.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 128
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 233
Illegal BglII site found at 1296
Illegal XhoI site found at 571 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 386
Illegal NgoMIV site found at 560
Illegal NgoMIV site found at 877
Illegal NgoMIV site found at 938 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 921
Illegal BsaI.rc site found at 421
Illegal BsaI.rc site found at 1395
2021 SZPT-China
Biology
As a photoactivated diguanylate cyclase (DGC), the chimeric protein BphS consists of the N-terminal photosensitive module of the BphG protein from Rhodobacter sphaeroides and the introduction of the R587A mutation into the RXXD motif from the C-terminal GGDEF domain of Slr1143. BphS protein is activated by changes in protein conformation under NIR light irradiation, thereby synthesizing c-di-GMP.
Usage
The coding sequences for BphS and BphO were inserted into the expression vector with BBa_K880005 (BBa_J23100 &BBa_B0034) to obtain J23100-B0034-bphS-pET RBS-bphO-rrnB T1 (BBa_K3740047). 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.