Toxicological and molecular characterization of pyrethroid-resistant horn flies, Haematobia irritans:: Identification of kdr and super-kdr point mutations
Toxicological and molecular characterization of pyrethroid-resistant horn flies, Haematobia irritans:: Identification of kdr and super-kdr point mutations
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DOI：
10.1016/s0965-1748(97)00057-x
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发表时间：
1997-08-01
影响因子：
3.8
通讯作者：
Kunz, SE
中科院分区：
文献类型：
--
作者：
Guerrero, FD;Jamroz, RC;Kunz, SE
两个拟除虫菊酯抗性品系对氯氰菊酯的抗性分别是敏感对照品系的17倍和688倍，对氯氟氰菊酯的抗性分别是敏感对照品系的17倍和11,300倍。与胡椒基丁醚的协同作用实验表明，超级抗性品系同时存在靶点不敏感和代谢抗性机制。利用逆转录聚合酶链式反应(RT-PCR)，克隆并测序了敏感和抗性家蝇钠通道基因的0.9kb片段，检测到两个序列变异，可能来自转录片段的选择性剪接。从抗性和感病家蝇基因片段推导的氨基酸序列是相同的，除了3个氨基酸替换，其中2个与家蝇的抗性有关，在两个抗性品系中都发现了与击倒抗性相关的亮氨酸对苯丙氨酸的替换(Kdr)。在超级抗性品系中发现蛋氨酸到苏氨酸的取代与超级kdr相关，Poly(A)(+)RNA的翻译和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)检测到翻译产物的浓度随着拟除虫菊酯抗性的增加而增加，超过260个DNA寡聚体的基因组DNA随机扩增多态性DNA(RAPD)-PCR产生一个与抗性相关的标记，命名为HF-77，该标记在任何敏感果蝇中都没有检测到，但在16%的抗性个体中存在，(C)1997 Elsevier Science Ltd.，所有权利保留。
Two pyrethroid-resistant strains of horn flies were found to be 17- and 688-fold more resistant to permethrin and 17- and 11,300-fold more resistant to cyhalothrin than a susceptible control strain, Synergism experiments with piperonyl butoxide showed that both target site insensitivity and metabolic resistance mechanisms were present in the Super Resistant strain, Using the reverse transcriptase-polymerase chain reaction (RT-PCR), a 0.9 kb fragment of the putative sodium channel gene from susceptible and resistant flies was cloned and sequenced, Two sequence variants were detected, presumably arising from alternative splicing of transcripts, The amino acid sequences deduced from the resistant and susceptible fly gene fragments were identical except for three amino acid substitutions, two of which have been associated with resistance in house flies, A leucine to phenylalanine substitution associated with knockdown resistance (kdr) was found in both resistant strains. A methionine to threonine substitution associated with super-kdr was found in the Super Resistant strain, Translation of poly(A)(+) RNA followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) detected translation products whose concentrations increased in association with pyrethroid resistance, Random-amplified polymorphic DNA (RAPD)-PCR of genomic DNA with over 260 DNA oligomers yielded one resistance-associated marker, designated HF-77, which was not detected in any susceptible flies but was present in 16% of the resistant individuals, (C) 1997 Elsevier Science Ltd, All rights reserved.