Composite

Part:BBa_K4286907:Design

Designed by: Bingru Feng   Group: iGEM22_SZU-China   (2022-09-28)


Suicide switch (Pcbh1-cbh1ls-mazEF-Tcbh1) for Trichoderma spp


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1006
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2663
    Illegal AgeI site found at 2007
    Illegal AgeI site found at 2707
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2644
    Illegal BsaI.rc site found at 2510
    Illegal BsaI.rc site found at 2544
    Illegal SapI site found at 2301


Design Notes

The Pcdh1-cdh1ls-mazEF-Tcdh1 circuit was integrated into pCAMBIA1302 vector by homologous recombination. We transformed the recombinant plasmid into Trichoderma atroviride by Agrobacterium-mediated transformation in order to achieve condition-triggered suicide of T. atroviride ultimately.


Source

The sequence of cbh1 promoter and cbh1 leader sequence belong to Trichoderma koningii cbh1 gene for 1,4-beta-D-glucan-cellobiohydrolase (GenBank: X69976.1), which comes from genomic sequence of Trichoderma koningii. The sequence of cbh1 terminator comes from genomic sequence of Trichoderma reesei.


References

[1]Kluge J, Terfehr D, Kück U. Inducible promoters and functional genomic approaches for the genetic engineering of filamentous fungi. Appl Microbiol Biotechnol. 2018 Aug;102(15):6357-6372. doi: 10.1007/s00253-018-9115-1.

[2]Wey TT, Hseu TH, Huang L. Molecular cloning and sequence analysis of the cellobiohydrolase I gene from Trichoderma koningii G-39. Curr Microbiol. 1994 Jan;28(1):31-9. doi: 10.1007/BF01575983.

[3]Madhavan A, Sukumaran RK. Promoter and signal sequence from filamentous fungus can drive recombinant protein production in the yeast Kluyveromyces lactis. Bioresour Technol. 2014 Aug;165:302-8. doi: 10.1016/j.biortech.2014.03.002. Epub 2014 Mar 12.

[4]Henrique-Silva F, el-Gogary S, Carle-Urioste JC, Matheucci E Jr, Crivellaro O, el-Dorry H. Two regulatory regions controlling basal and cellulose-induced expression of the gene encoding cellobiohydrolase I of Trichoderma reesei are adjacent to its TATA box. Biochem Biophys Res Commun. 1996 Nov 12;228(2):229-37. doi: 10.1006/bbrc.1996.1646.

[5]Zhong Y, Liu X, Xiao P, Wei S, Wang T. Expression and secretion of the human erythropoietin using an optimized cbh1 promoter and the native CBH I signal sequence in the industrial fungus Trichoderma reesei. Appl Biochem Biotechnol. 2011 Nov;165(5-6):1169-77. doi: 10.1007/s12010-011-9334-8. Epub 2011 Aug 16.

[6]Karhunen T, Mäntylä A, Nevalainen KM, Suominen PL. High frequency one-step gene replacement in Trichoderma reesei. I. Endoglucanase I overproduction. Mol Gen Genet. 1993 Dec;241(5-6):515-22. doi: 10.1007/BF00279893.