Device

Part:BBa_K3740030

Designed by: Guiyi Huang   Group: iGEM21_SZPT-CHINA   (2021-09-20)


J23100-B0034-bphS-pET RBS-bphO-rrnB T1-J23109-B0034-fcsR-rrnB T1

Description

This is a device that is used to regulate the level 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 NheI site found at 2818
    Illegal NheI site found at 2841
    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
    Illegal XhoI site found at 3577
  • 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 NgoMIV site found at 2935
    Illegal NgoMIV site found at 3680
    Illegal AgeI site found at 2634
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 982
    Illegal BsaI site found at 3574
    Illegal BsaI.rc site found at 482
    Illegal BsaI.rc site found at 1456
    Illegal BsaI.rc site found at 3322
    Illegal SapI.rc site found at 3687


2021 SZPT-China

Biology

This is a device that is used to regulate the level of c-di-GMP.

Usage

We connected J23100-B0034-bphS-pET RBS-bphO-rrnB T1 (BBa_K3740047) and J23109-B0034-fcsR-rrnB T1 (BBa_K3740048) to the expression vector pSEVA331 through the expression vector pSEVA331. Then the ligated mixture was introduced into E. coli DH5α for replication, and then the plasmid was transferred into G. hansenii ATCC 53582, so that G. hansenii ATCC 53582 could control BC production through light regulation.

Figure 1. Gene circuit of J23100-B0034-bphS-pET RBS-bphO-J23109-B0034-fcsR-rrB T1

Characterization

1. Identification

As shown in Figure 2, composite part BBa_K3740030 was identified successfully by PCR amplification.

Figure 2. Agarose gel image of J23100-B0034-bphS-pET RBS-bphO-rrnB T1-J23109-B0034-fcsR-rrnB T1. BBa_K3740030, 3765bp.

2.Characterization

A significant difference in BC film production under NIR light and dark conditions was observed for strain, J23100-B0034-bphS-pET RBS-bphO-J23109-B0034-fcsR-rrB T1-pSEVA331-G. hansenii ATCC 53582, indicating that G. hansenii ATCC 53582 could control BC production through light regulation.

Figure 3. BC film production by different strains. (a) J23100-B0034-bphS-pET RBS-bphO-J23109-B0034-fcsR- rrnB T1-pSEVA331-G. hansenii ATCC 53582; (b) pSEVA331-G. hansenii ATCC 53582.

References

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

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Categories
Parameters
None