关键词: cry1A基因, 简并引物, 方法建立, 成分检测, 转基因监管

Abstract: cry1A gene encodes Bacillus thuringiensis insecticidal crystal protein, which is the most widely used insectresistant gene in genetically modified (GM) plants, including cry1Ab, cry1Ac, cry1A.105, cry1Ab/Ac, etc.. The detection method on targeting cry1A gene has particularly important in GM plants safety supervision and detection. Several cry1A gene detection method has been developed, but because of much difference among this gene sequences, the detection method can’t cover major GM events. Thus, this paper adopted the strategy of degenerate primers to develop a universal detection method for cry1A gene, detection capability covered 16 GM events. The development process included the establishment of detection method in the internal laboratory and the collaborative validation between laboratories. The results showed that this method could specifically detect the cry1A gene in the samples and limit of detection (LOD) was 01% under the condition of 04 μmol·L-1 primer concentration and 64 ℃ annealing temperature. The detection method of cry1A gene had comprehensive parameters, good repeatability and reproducibility, which providing a method reference of GMO supervision and detection.

Key words: cry1A gene, degenerate primer, method development, ingredient detection, GMO supervision