A vital staining method for measuring the efficiency of transfection of eucaryotic cells.
The xylE gene encodes catechol 2,3-dioxygenase, which catalyzes the conversion of catechol to 2-hydroxymuconic semialdehyde. The expression of this gene in eucaryotic cells can be detected simply by addition of catechol to the growth medium of the cells: cells that have a sufficient level of expression of the xylE gene stain yellow because of the accumulation of 2-hydroxymuconic semialdehyde. The number of stained cells is thus dependent upon the transfection efficiency as well as the level of expression of the xylE gene and is a measure of the combined transfection/expression efficiency in a particular cell type. Since the staining procedure does not affect the viability of the culture, the cells can be harvested afterward and analyzed for the expression of other, cotransfected, genes. This system for measuring transfection efficiency is especially useful when only small amounts of tissue are available.
D000818:Animals; D050557:Catechol 2,3-Dioxygenase; D002396:Catechols / Q000378:metabolism; D002642:Chick Embryo; D003116:Color; D049308:Dioxygenases; D005231:Fatty Acids, Unsaturated / Q000032:analysis; D010105:Oxygenases / Q000032:analysis / Q000235:genetics*; D010957:Plasmids; D013194:Staining and Labeling; D014162:Transfection
D002396:Catechols; D005231:Fatty Acids, Unsaturated; C002091:hydroxymuconic semialdehyde; D010105:Oxygenases; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase; C034221:catechol