Herbicidal addition assays

Site-directed mutagenesis was used to make additional amino acid substitutions at these sites in yeast ALS. At some of the sites, e.g. alall7, prol92, or trp586, nearly any substitution for the wild type amino acid that was tested resulted in a herbicide-resistant enzyme (Table I). Each of the mutant enzymes was characterized by enzyme assays to compare its activity, and its sensitivity to the sulfonylurea herbicide chlorimuron ethyl, to the wild type enzyme. These analyses have indicated that some of the mutations have little adverse effect on the activity of the enzyme, while decreasing sensitivity to the herbicide from three to greater than one thousandfold. The characteristics of these mutant enzymes were further evaluated in vivo in order to investigate the utility of particular herbicide/mutant enzyme combinations (Falco et al., manuscript in preparation). 

The evaluation of a number of immunoassay diagnostic kits was undertaken to determine their usefulness in a regulatory analytical laboratory environment in the food, feed and pesticide areas. Four rapid enzyme immunoassay tests for the detection of aflatoxin residues at the 20 ppb level in animal feeds were compared to the official HPLC procedure. In the pesticide area, a commercial pentachlorophenol competitive inhibition assay for residues in water was investigated as to its applicability to poultry and pork liver matrices. In addition, an ELISA screening procedure for the herbicide fusilade was developed. Modifications were incorporated into the rapid immunoband 1-2 Test procedure for the detection of motile Salmonella in various food and animal feed products resulting in quicker analysis than the standard culture method. Also, a comparative evaluation of a Quik-Card Test for sulphamethazine drug residues in pork urine, liver and muscle tissue, is described. 

Fig. 33.6. Supplementation assay of Lenina gibba plants Plants grow normally in water without Compound A (left). Addition of Compound A prevents plants from growing (middle). Addition of the three branched-chain amino acids valine (Val), leucine (Leu) and isoieucine (lie) overcomes the herbicidal effect of Compound A and restores piant growth (right).

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