Coding
PC2

Part:BBa_K3979008:Design

Designed by: Akshay J, Abhishek Raghunathan   Group: iGEM21_IISER_TVM   (2021-10-18)

Plant Chitinase Combo 2


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 7
    Illegal XhoI site found at 1030
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This is a chitinase gene designed using the catalytic domain(CD) of Triticum aestivum cultivar Sumai 3 and the chitin-binding domain (CBD) of Hordeum vulgare cultivar NK1558. We created a chimeric chitinase with a wheat catalytic domain flanked by two barley chitin-binding domains. The protein sequences were linked initially and then reverse translated to nucleotide sequences(stop codon inclusive) along with codon optimization for E.coli expression using the BOOST Juggler tool. BamHI and HindIII sites were added at the 5’ and 3’ end of the gene, respectively.


Source

Chitinase II precursor from Triticum aestivum GenBank ID: AAD28730.1

Chitinase from Hordeum vulgare GenBank ID: AAA18586.1


References


  • Singh A, Kirubakaran SI, Sakthivel N. Heterologous expression of new antifungal chitinase from wheat. Protein Expr Purif. 2007 Nov;56(1):100-9. doi: 10.1016/j.pep.2007.06.013. Epub 2007 Jul 12. PMID: 17697785.
  • Kirubakaran SI, Sakthivel N. Cloning and overexpression of antifungal barley chitinase gene in Escherichia coli. Protein Expr Purif. 2007 Mar;52(1):159-66. doi: 10.1016/j.pep.2006.08.012. Epub 2006 Sep 6. PMID: 17029984.
  • O. Trott, A. J. Olson, AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization and multithreading, Journal of Computational Chemistry 31 (2010) 455-461 DOI 10.1002/jcc.21334