淡色库蚊抗性种群和敏感品系中铁蛋白基因表达量的分析

Comparative analysis on expression levels of the ferritin gene in Culex pipiens pallens

目的对淡色库蚊抗性种群和敏感品系中的铁蛋白基因表达量进行分析。方法设计引物,扩增目的基因全长、纯化测序、序列分析,荧光定量PCR检测铁蛋白基因的表达水平;采用假设检验中的t检验方法,对2种不同处理的淡色库蚊样本体内铁蛋白基因表达量进行统计分析。结果成功扩增淡色库蚊铁蛋白基因片段,经荧光定量PCR测定,淡色库蚊敏感品系铁蛋白荧光定量(F敏)和抗性种群铁蛋白荧光定量(F抗)平均值分别为11.41和15.73,内参基因荧光定量(β敏和β抗)平均值分别为10.03和18.27,铁蛋白基因在淡色库蚊抗性种群中的表达量是敏感品系的15.08倍,t检验结果显示,两样本体内铁蛋白基因表达差异有统计学意义(t=26.100,P<0.001)。结论淡色库蚊铁蛋白基因可作为蚊虫抗性检测及治理的靶标基因。

ObjectiveTo analyze the ferritin gene expression levels in different strains of Culex pipiens pallens.MethodsThe primers were designed and synthesized, target gene was amplified by fluorescent quantitative PCR to detectthe expression level between resistant strain and susceptible strains of Cx. pipiens pallens. A t test method was used todetect the statistical significance for the two samples.ResultsThe ferritin gene of Cx. pipiens pallens was clonedsuccessfully. The quantitative average of ferritin fluorescence was 11.41 in susceptible strain and 15.73 in resistant strain;the quantitative average of reference gene fluorescence was 10.03 in susceptible strain, and 18.27 in resistant strain.Fluorescent quantitative PCR proved the ferritin gene expression level of resistant strain was 15.08 times that of susceptiblestrain, t test showed that two samples of ferritin gene expression differences were statistically significant（t=26.100, P〈0.001）.Conclusion Culex pipiens pallensferritin gene can be used as a new gene target for resistant detection and management.