EVALUATION OF A COMMERCIAL ELISA KIT FOR QUANTIFYING CRY1AC PRODUCTION IN GLANDLESS COTTON

摘要

Genes from Bacillus thuringiensis (Bt) have been genetically engineered into cotton (Gossypium spp.) plants to promote insect resistance. For this study, we were interested in quantifying the expression of the Cry1Ac protein encoded by an introduced Bt gene. The amount of Cry1Ac protein produced by plants in four cotton backcross populations—segregating for the glandless gene (Gl_2~e )—and their respective recurrent Bt-containing parents was quantified using the ELISA-based Envirologix QuantiPlateKit for Cry1Ab/Cry1Ac. Our objective was to use this kit to identify glandless plants with two copies of the Bt gene (homozygous) versus glandless plants with one copy (heterozygous). Our results clearly indicated the lack of Cry1Ac protein in our negative control while showing a continuous range of values for those plants expressing Cry1Ac. There was no definite break among the values that would indicate heterozygotes versus homozygotes having twice the expression level of the heterozygotes. Although we were unable to identify homozygotes with this kit, the kit was easy to use and gave a range of values to select plants with the highest Cry1Ac expression.