论文部分内容阅读
目的 制备敌敌畏抗性品系淡色库蚊酯酶B1、A2探针 ,并应用于抗性品系及敏感品系的检测。 方法 分别设计酯酶B1、A2上下游引物 ,5’端地高辛标记 ,PCR法制备DNA探针 ,分子杂交检测各个品系淡色库蚊。 结果 摸索出合适的区分抗性与敏感品系的探针浓度 ,在该浓度下各品系杂交点的阳性率有差别。应用酯酶B1探针检测各品系蚊虫 ,抗敌敌畏品系的阳性率为 40 .0 % ,抗残杀威品系的阳性率为 2 3 .3 % ,抗氯氰菊酯品系的阳性率为 18 3 % ,敏感品系的阳性率为 11 7% ;应用酯酶A2探针检测各品系蚊虫 ,抗敌敌畏品系的阳性率为 43 3 % ,抗残杀威品系的阳性率为 2 5 0 % ,抗氯氰菊酯品系的阳性率为 2 1 7% ,敏感品系的阳性率为 15 0 %。 结论 应用酯酶B1、A2探针能够区分抗性品系 ,因此能够用于抗性的测定。从而为抗有机磷类杀虫剂媒介昆虫基因检测的实用性和抗性分子生物学机制的研究打下基础。
OBJECTIVE To prepare the dichlorvos resistant Esterase B1 and A2 probe of Culex pipiens quinquefasciatus and to detect the resistant and susceptible strains. Methods Esterase B1, A2 upstream and downstream primers were designed respectively. Digoxigenin was labeled on the 5 ’end. DNA probes were prepared by PCR, and Culex pipiens pallens were tested by molecular hybridization. The results obtained a suitable probe concentration to distinguish between resistant and susceptible strains, at which the positive rate of hybridization points in each strain varied. The esterase B1 probe was used to detect the mosquitoes in each strain. The positive rate of anti-dichlorvos strain was 40.0%, the positive rate of anti-malachite strain was 23.3%, the positive rate of anti-cypermethrin strain was 18.3% The positive rate was 11 7%. The esophageal A2 probe was used to detect the mosquitoes in each strain. The positive rate of anti-dichlorvos strain was 43.3% and that of anti-marcant strain was 25.0%. The positive rate of resistance to cypermethrin strain was 21.7%, the positive rate of sensitive strains was 15 0%. Conclusion Esterase B1 and A2 probes can be used for the determination of resistance in resistant strains. Thus laying a foundation for the study of the practicality and molecular mechanism of resistance to organophosphate insecticide gene detection.