Coding

Part:BBa_K1470001

Designed by: Pascal Sartor   Group: iGEM14_Freiburg   (2014-10-01)
Revision as of 21:05, 1 October 2014 by Suricate (Talk | contribs)

puromycin N-acetyl-transferase

Puromycin is an antibodic derived from the bacteria Streptomyces alboniger. The substance affects the cell both in protein translation and mitochondrial protein import [1]. Due to this, it's toxic to many organisms such as bacteria, Trypanosoma and humans.
A small part of its structure resembles to aminoacetylated tRNA and is recognized by ribosomes. The nascent chain containing puromycin leads to a translational stop.
Streptomyces alboniger protects itself from puromycin with an acetyl transferase. It uses acetyl-CoA to bind the acetyl moiety to the amino group. In this state puromycin cannot be recognized by the ribosom anymore [2].
This enzyme is often used for selecting stable eucaryotic cell lines. Because of the blocked protein biosynthesis more than 99 % of uninfected cells will be dead within two days.

References

[1] NEUBERT, D. AND LEHNINGER, A. L.: The effect of oligomycin, gramicidin, and other antibiotics on reversal of mitochondrial swelling by adenosine triphosphate. Biochim. biophys. Acta 62: 556-565, 1962.
[2] DARKEN MA.: PUROMYCIN INHIBITION OF PROTEIN SYNTHESIS. Pharmacol Rev. 1964 Sep;16:223-43.
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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 481


[edit]
Categories
Parameters
None