Functional Reconstitution and Biophysical Characterization of a Putative Caenorhabditis Elegans K+ Channel

Abstract

The focus of this project was to pharmacologically and biochemically characterize a CeK (F17c85) gene product through the expression of its cRNA in the Xenopus laevis system. CeK is believed to be a structural homolog to two K+ channel genes, TWIK-1 and TOK-l. The clone was amplified from a Caenorhabditis elegans cDNA library and was transformed into a pCR 2.1, an E. coli vector. The plasmid was then able to be transcribed into cRNA for injection into oocytes from Xenopus laevis. After injection, oocytes were analyzed using 2-microelectrode patch clamping techniques which measure the electrical potential across the cell. The functional expression of CeK was determined using a voltage clamping technique to measure the ionic chemical activity across the oocyte. The insertion of CeK cRNA resulted in little or no expression of a functional K+ channel in the oocyte system.