Part:BBa_K3740047
J23100-B0034-bphS-pET RBS-bphO-rrnB T1
Description
Near-infrared light (NIR light) activated synthesis of c-di-GMP in Gluconacetobacter hansenii ATCC 53582 and synthesis of c-di-GMP.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NotI site found at 189 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 294
Illegal BglII site found at 1357
Illegal XhoI site found at 632
Illegal XhoI site found at 2489 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 447
Illegal NgoMIV site found at 621
Illegal NgoMIV site found at 938
Illegal NgoMIV site found at 999
Illegal NgoMIV site found at 2678
Illegal AgeI site found at 2634 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 982
Illegal BsaI.rc site found at 482
Illegal BsaI.rc site found at 1456
2021 SZPT-China
Biology
This is a combination of bphS (BBa_K3740019) and bphO (BBa_K3740001) to form a generator that is used to regulate the level of c-di-GMP.
Usage
The encoding 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.
Characterization
1. Identification
As shown in Figure 3, composite part BBa_K3740047 was identified successfully by PCR amplification.
2. Characterization
As shown in Figure 4, J23100-bphS-pET RBS-bphO-rrnB T1-pSEVA331-G. hansenii ATCC 53582, produced a higher level of BC film under NIR light than that under dark conditions, indicating that BphS in G. hansenii ATCC 53582 can synthesize c-di-GMP.
References
[1] Min-Hyung, Gomelsky, Mark. Near-infrared Light Responsive Synthetic c-di-GMP Module for Optogenetic Applications.
| None |