Difference between revisions of "Part:BBa K3740047"

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<partinfo>BBa_K3740047 short</partinfo>
 
<partinfo>BBa_K3740047 short</partinfo>
 
===Description===
 
===Description===
−
Near-infrared light activated synthesis of c-di-GMP in <i>Gluconacetobacter hansenii ATCC 53582</i> and synthesis of c-di-GMP.
+
Near-infrared light activated synthesis of c-di-GMP in <i>Gluconacetobacter hansenii</i> ATCC 53582 and synthesis of c-di-GMP.
  
 
===Sequence and Features===
 
===Sequence and Features===

Revision as of 14:37, 18 October 2021


J23100-B0034-bphS-pET RBS-bphO-rrnB T1

Description

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

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NotI site found at 189
  • 21
    INCOMPATIBLE 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
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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.

Figure 2. Gene circuit of J23100-B0034-bphS-pET RBS-bphO-rrnB T1

Characterization

1. Identification

As shown in Figure 3, composite part BBa_K3740047 was identified successfully by PCR amplification.

Figure 3. Agarose gel electrophoresis image of J23100-B0034-bphS-pET RBS-bphO-rrnB T1. BBa_K3740047, 2782bp

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 near-infrared light than that under dark conditions, indicating that BphS in G. hansenii ATCC 53582 can synthesize c-di-GMP.

Figure 4. BC yield by J23100-bphS-pET RBS-bphO-rrnB T1-pSEVA331-G. hansenii ATCC 53582 under NIR light illumination and dark conditions.

References

[1] Min-Hyung, Gomelsky, Mark. Near-infrared Light Responsive Synthetic c-di-GMP Module for Optogenetic Applications.