abstract |
The present invention relates to methods of producing, isolating, identifying and characterizing herbicide tolerant ACCase variants. In one embodiment, the invention encompasses a method of screening for an acetyle-CoA carboxylase (ACCase) enzyme which is tolerant to at least one herbicide, comprising: a) providing an ACCase-deficient yeast with a chimeric ACCase, said chimeric ACCase comprising: at least two regions that further comprise: i) an N-terminal region, said N-terminal region derived from yeast, fungi or monocot cytoplasmic ACCases, preferably a yeast ACCase; ii) a C-terminal region, said C-terminal region derived from monocot plastific ACCases and comprising an HSR; and iii) said N-terminal region comprises about 50% to about 60% of the chimeric ACCase; b) isolating herbicide tolerant yeast cells after culturing in the presence of at least one herbicide; and c) further comprising identifying the mutation(s) not present in chimeric ACCase prior to culturing, which confers tolerance to at least one herbicide. |