Coding

Part:BBa_K3740019

Designed by: Guiyi Huang   Group: iGEM21_SZPT-CHINA   (2021-08-26)
Revision as of 13:20, 18 October 2021 by HUANG (Talk | contribs) (2021 SZPT-China)


bphS, photo-activated diguanylate cyclase

Description

Near-infrared light activated synthesis of c-di-GMP in Gluconacetobacter hansenii ATCC 53582.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 128
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 233
    Illegal BglII site found at 1296
    Illegal XhoI site found at 571
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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.

Figure 1. Engineering a potent photoactivated DGC

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-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.

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