Genetic screening for functionality of bacterial potassium channel mutants using K+ uptake-deficient Escherichia coli.
Potassium channels play an essential role in a wide range of biological processes, including cell volume regulation and the maintenance and control of electrical signals. With the advent of the structural era of ion channel biology, it has become critical to learn more about the functional properties of the prokaryotic channels, and this is the area in which genetic screens have become an increasingly useful approach. Here, we describe a bacteria-based complementation assay that we applied to investigate gating mutants of the prokaryotic K+ channel MthK, which was cloned from the archeon Methanobacterium thermoautotrophicum. The results demonstrated that heterologously expressed MthK is fully assembled and functional in Escherichia coli. This complementation assay should be useful in the initial identification of prokaryotic K+ channel mutants that result in altered channel function.
Parfenova
Lyubov V
LV
Department of Physiology, University of Texas Health Science Center, San Antonio, USA.
Rothberg
Brad S
BS
D015153:Blotting, Western; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D005798:Genes, Bacterial / Q000235:genetics; D005816:Genetic Complementation Test; D017136:Ion Transport; D017014:Methanobacterium / Q000737:chemistry; D009154:Mutation / Q000235:genetics*; D011188:Potassium / Q000378:metabolism*; D015221:Potassium Channels / Q000235:genetics* / Q000378:metabolism*; D014169:Transformation, Bacterial
D003470:Culture Media; D015221:Potassium Channels; D011188:Potassium