Composite

Part:BBa_K842015

Designed by: Sean P. Curran, Percy Genyk, Ellen Park, Rebecca, Gao, Stephan Genyk, Megan Bernstein, Rachel Kohan, Eric Siryj, Luke Quinto   Group: iGEM12_USC   (2012-10-03)
Revision as of 15:43, 10 October 2012 by Seancurran (Talk | contribs)

T7 RBS cheY

USC T7 RBS cheY.gif

T7 serves as a promoter that is stimulated by IPTG. RBS is a sequence in DNA located upstream of the start codon. It affects the rate at which the open reading frame is translated. CheY is regulatory protein that reverses the direction of flagella rotation from counter-clockwise to clockwise. When the methyl-accepting chemotaxis protein (MCP) is not bound to any attractant, cheA is activated and told to self-phosphorylate itself by hydrolyzing ATP. Once the cheA is phosphorylated, it transfers its phosphate group to cheY, therefore activating it. CheY then moves to the cytoplasmic side of the flagella apparatus and binds to it. When flagella are not bound to cheY, it rotates in a counter-clockwise direction, which allows the bacteria to move in a “forward” direction. Flagella that are bound to phosphorylated cheY switches its rotation to clockwise, which induces tumbling. Used for researcher control of signaling mechanisms that influence flagella function.

Cloning and Uses This part was generated via PCR from the E. coli strain DH5α and cloned into the vector pSB1C3. It is to be used with other chemotaxis genes to form a complete regulatory pathway that controls the response system in MCP and the direction of rotation in flagella. http://uscigem2012.files.wordpress.com/2012/10/cheytumbling.gif

Sequence and Features


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
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None