Field L M, Crick S E, Devonshire A L
BEC Department, IACR-Rothamsted, Harpenden, Herts.
Insect Mol Biol. 1996 Aug;5(3):197-202. doi: 10.1111/j.1365-2583.1996.tb00054.x.
The ability of a peach-potato aphid (Myzus persicae) to resist insecticides can depend on both the presence of amplified esterase genes and on differences in their expression. Here we report a polymerase chain reaction-restriction enzyme (PCR-REN) technique which can detect the presence of amplified esterase genes and distinguish between the two possible types of amplified gene (E4 and FE4) and a homologous fragment from susceptible aphids. The technique is quick and sensitive enough to be used on a fraction of an aphid or an individual embryo. Furthermore, it can be combined with a pre-PCR digestion using a methylation-sensitive enzyme (Hpall) to determine whether or not the esterase genes contain 5-methylcytosine, the presence or absence of which is known to correlate with changes in gene expression.
桃蚜（Myzus persicae）对杀虫剂的抗性能力既取决于扩增酯酶基因的存在，也取决于其表达差异。在此，我们报告一种聚合酶链反应-限制性酶切（PCR-REN）技术，该技术能够检测扩增酯酶基因的存在，并区分两种可能的扩增基因类型（E4和FE4）以及来自敏感蚜虫的同源片段。该技术快速且灵敏，足以用于蚜虫的一部分或单个胚胎。此外，它可以与使用甲基化敏感酶（HpaII）的PCR前消化相结合，以确定酯酶基因是否含有5-甲基胞嘧啶，已知其存在与否与基因表达变化相关。
