Talk:CH391L/S13/GeneticMarkers: Difference between revisions

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*'''[[User:Gabriel Wu|Gabriel Wu]] 17:33, 4 March 2013 (EST)''': For those who work outside of E. coli, there's a class of selection markers used more commonly in yeast known as auxotrophic markers. Maybe not this time, but this topic should be addressed on this page at some point.
*'''[[User:Gabriel Wu|Gabriel Wu]] 17:33, 4 March 2013 (EST)''': For those who work outside of E. coli, there's a class of selection markers used more commonly in yeast known as auxotrophic markers. Maybe not this time, but this topic should be addressed on this page at some point.
**'''[[User:Benjamin Gilman|Benjamin Gilman]] 17:39, 5 March 2013 (EST)''': The [http://openwetware.org/wiki/CH391L/S12/Selectablegeneticmarkers selectable genetic markers] page from last year's class gives a brief overview of different categories of markers, including auxotrophic markers.  Briefly, auxotrophs are organisms that are incapable of synthesizing a particular molecule required for growth, in this case because they've had a necessary gene knocked out.  The mutant auxotrophs grow when supplemented with the required molecule but require complementation with the deleted gene to survive without it .  Auxotrophic markers are used more often in yeast than bacteria mostly because the range of antibiotic resistance markers in yeast is much smaller.  The GFAT-expressing gene I discussed in class is an example of an auxotrophic marker which works in ''E. coli'', and a related GFAT gene works similarly as a marker in fission yeast (''S. pombe'').

Revision as of 15:39, 5 March 2013

  • Gabriel Wu 17:30, 4 March 2013 (EST): On a practical note, ampicillin (amp), kanamycin (kan), and chloramphenicol (cam) are the most used antibiotic selection markers in synthetic biology. Amp in my experience is convenient, but somewhat "fragile" (relative to kan and cam). Convenient: typically "rescuing" your cells post-transformation is unnecessary when using amp. "Rescuing" is necessary for kan and cam. Cam and kan are useful for long time course experiments (longer than 24 hours) because amp generally breaks down (or is broken down) over time.
  • Gabriel Wu 17:33, 4 March 2013 (EST): For those who work outside of E. coli, there's a class of selection markers used more commonly in yeast known as auxotrophic markers. Maybe not this time, but this topic should be addressed on this page at some point.
    • Benjamin Gilman 17:39, 5 March 2013 (EST): The selectable genetic markers page from last year's class gives a brief overview of different categories of markers, including auxotrophic markers. Briefly, auxotrophs are organisms that are incapable of synthesizing a particular molecule required for growth, in this case because they've had a necessary gene knocked out. The mutant auxotrophs grow when supplemented with the required molecule but require complementation with the deleted gene to survive without it . Auxotrophic markers are used more often in yeast than bacteria mostly because the range of antibiotic resistance markers in yeast is much smaller. The GFAT-expressing gene I discussed in class is an example of an auxotrophic marker which works in E. coli, and a related GFAT gene works similarly as a marker in fission yeast (S. pombe).