Difference between revisions of "Part:BBa K3740019"
 
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<partinfo>BBa_K3740019 short</partinfo>
 
<partinfo>BBa_K3740019 short</partinfo>
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===Description===
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Near Infrared light(NIR) activated synthesis of c-di-GMP in <i>Gluconacetobacter hansenii</i> ATCC 53582.
  
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Description
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===Sequence and Features===
  
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Near-infrared light activated synthesis of c-di-GMP in Acetobacter xylinum
 
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===Usage and Biology===
 
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<span class='h3bb'>Sequence and Features</span>
 
 
<partinfo>BBa_K3740019 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3740019 SequenceAndFeatures</partinfo>
  
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<partinfo>BBa_K3740019 parameters</partinfo>
 
<partinfo>BBa_K3740019 parameters</partinfo>
 
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=2021 SZPT-China=
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<h3>Biology</h3>
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<p>As a photoactivated diguanylate cyclase (DGC), the chimeric protein BphS consists of the N-terminal photosensitive module of the BphG protein from <i>Rhodobacter sphaeroides</i> 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.</p>
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[[File:szpt18.png|400px|thumb|center|Figure 1. Engineering a potent photoactivated DGC]]
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<h3>Usage</h3>
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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.
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[[File:Szpt11.png|400px|thumb|center|Figure 2. Gene circuit of bphS]]

Latest revision as of 16:40, 18 October 2021


bphS, photo-activated diguanylate cyclase

Description

Near Infrared light(NIR) 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-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.

Figure 2. Gene circuit of bphS