Difference between revisions of "Part:BBa K1680007"
 
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===(Characterized by iGEM TU Darmstadt-2020)<br>===  
 
===(Characterized by iGEM TU Darmstadt-2020)<br>===  
  
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We conducted some further literature research in order to contribute to further application of the Cre recombinase. The Cre recombinase has a molecular weight of 38 kDA, needs no cofactors and has its optimal reaction temperature at 37°C and above[1]. Most likely a certain threshold level of Cre activity has to be reached, in order to achieve high recombination efficiency[2]. Recombination is achieved by the binding of four recombinase monomers to each palindromic site of the loxP-site, the specific recombination target sites of the Cre recombinase, forming a tetrameric enzyme complex. Upon complex formation, recombination sites are brought together and the exchange of DNA flanked by loxP-sites is induced.[3]  
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We conducted some further literature research in order to contribute to further application of the Cre recombinase. The Cre recombinase has a molecular weight of 38 kDA, needs no cofactors and has its optimal reaction temperature at 37°C and above[1]. Most likely a certain threshold level of Cre activity has to be reached, in order to achieve high recombination efficiency[2]. Recombination is achieved by the binding of four recombinase monomers to the palindromic sites of the loxP-sites, the specific recombination target sites of the Cre recombinase, forming a tetrameric enzyme complex. Upon complex formation, recombination sites are brought together and the exchange of DNA flanked by loxP-sites is induced.[3]  
  
 
   
 
   

Latest revision as of 09:46, 22 October 2020

Cre recombinase

This part contains the protein coding sequence for the DNA recombinase Cre and is codon optimized for yeast. Cre recombinases recognize two loxP sites (BBa_K1680005) and depending on their orientation cuts out or reverses the DNA sequence between them.


Sequence and Features

This part is submitted in pSB1C3 with RFC25 standard. The part only sequence does therefore not contain a start and stop codon.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 437


General Information for Cre recombinase

(Characterized by iGEM TU Darmstadt-2020)

We conducted some further literature research in order to contribute to further application of the Cre recombinase. The Cre recombinase has a molecular weight of 38 kDA, needs no cofactors and has its optimal reaction temperature at 37°C and above[1]. Most likely a certain threshold level of Cre activity has to be reached, in order to achieve high recombination efficiency[2]. Recombination is achieved by the binding of four recombinase monomers to the palindromic sites of the loxP-sites, the specific recombination target sites of the Cre recombinase, forming a tetrameric enzyme complex. Upon complex formation, recombination sites are brought together and the exchange of DNA flanked by loxP-sites is induced.[3]


Reference

[1]: [Frank Buchholz, Leonie Ringrose, Pierre-Olivier Angrand, Fabio Rossi, A. Francis Stewart, Different Thermostabilities of FLP and Cre Recombinases: Implications for Applied Site-Specific Recombination, Nucleic Acids Research, Volume 24, Issue 21, 1 November 1996, Pages 4256–4262, https://doi.org/10.1093/nar/24.21.4256]  

[2]: Brian Sauer and Nancy Henderson, Site-specific DNA recombination in mammalian cells by recombinase of bacteriophage P1, Proc. Natl. Acad. Sci. USA, 1988, Vol. 85, pages 5166-5170, doi: 85 14 5166-5170 

[3] Andras Nagy et al., Cre Recombinase: The Universal Reagent for Genome Tailoring, Wiley-Liss Inc., 1999, genesis 26, pages 99-109, https://doi.org/10.1002/(SICI)1526-968X(200002)26:2<99::AID-GENE1>3.0.CO;2-B