School of Biological Sciences, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.
School of Biological Sciences, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.
Pestic Biochem Physiol. 2019 Jan;153:116-121. doi: 10.1016/j.pestbp.2018.11.011. Epub 2018 Nov 17.
Glutathione S-transferases (GSTs) from susceptible Aedes albopictus larvae were partially isolated using two different purification strategies (GSTrap™ HP and GSH-agarose affinity columns) and the effects of permethrin and DDT on expression of the GSTs were investigated. Distinct double bands on SDS-PAGE with molecular weights between 20 and 25 kDa were successfully purified using GSTrap™ HP while a single band of 24.5 kDa was purified using GSH-agarose. The isolated GSTs belonged to the Delta, Sigma and Theta GST classes. When exposed to permethrin, one isoform of Theta, four isoforms of Sigma and thirteen isoforms of Delta GSTs showed an increased expression between 1.4-fold and 2.5-fold while DDT treatment resulted in between 1.4-fold and 3.2-fold increased expression in one isoform of Theta, four isoforms of Sigma and eleven isoforms of Delta GSTs (p < .05). This study indicated that GSTrap™ HP was more competent in isolating Ae. albopictus GSTs compared to GSH-agarose and also variable expression of GST isoforms occur in response to different insecticides. This information may be useful for improving insecticide resistance management strategies in aspect of molecular resistant and evolutionary tolerant detoxification enzyme.
谷胱甘肽 S-转移酶(GSTs)从敏感的白纹伊蚊幼虫中被部分分离出来,使用了两种不同的纯化策略(GSTrap™ HP 和 GSH-琼脂糖亲和柱),并研究了拟除虫菊酯和滴滴涕对 GSTs 表达的影响。使用 GSTrap™ HP 成功地纯化了 SDS-PAGE 上分子量在 20 到 25 kDa 之间的两个明显的双带,而使用 GSH-琼脂糖纯化了 24.5 kDa 的单带。分离出的 GST 属于 Delta、Sigma 和 Theta GST 类。当接触拟除虫菊酯时,一个 Theta 同工酶、四个 Sigma 同工酶和十三个 Delta GST 同工酶的表达增加了 1.4 到 2.5 倍,而滴滴涕处理导致一个 Theta 同工酶、四个 Sigma 同工酶和十一个 Delta GST 同工酶的表达增加了 1.4 到 3.2 倍(p <.05)。这项研究表明,与 GSH-琼脂糖相比,GSTrap™ HP 更能有效地分离白纹伊蚊 GST,而且 GST 同工酶的表达也会因不同的杀虫剂而发生变化。这些信息可能有助于改进杀虫剂抗性管理策略,从分子抗性和进化耐受解毒酶方面考虑。