Serine proteinase over-expression in relation to deltamethrin resistance in Culex pipiens pallens
Serine proteinase over-expression in relation to deltamethrin resistance in Culex pipiens pallens
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DOI：
10.1016/j.abb.2005.03.021
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发表时间：
2005-06-01
影响因子：
3.9
通讯作者：
Zhu, CL
中科院分区：
文献类型：
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作者：
Gong, MQ;Shen, B;Zhu, CL
采用抑制减法杂交和大阵列基因表达谱分析相结合的方法，从淡色库蚊中分离出两个丝氨酸蛋白酶基因，作为抗溴氰菊酯菌株的显著上调基因。这两个基因在抗性品系中的表达量至少是敏感品系的三倍。利用cDNA末端快速扩增技术筛选构建的cDNA文库，克隆了这两个序列。胰蛋白酶cDNA （GenBank/NCBI AY034060）全长909 bp，开放阅读框786 bp，共261个氨基酸，与冈比亚按蚊的胰蛋白酶基因最为相似。乳糜蛋白酶cDNA （GenBank/NCBI AF468495）全长992 bp，开放阅读框816 bp，推导出的氨基酸序列有271个氨基酸，与埃及伊蚊乳糜蛋白酶样蛋白最相似。这两个基因在蚊子C6/36细胞中稳定表达，Western blot结果显示，两个基因的条带分别为29和30 kDa。在这些细胞中，经过溴氰菊酯处理后，它们对生存能力有保护作用。结果表明，胰酶和凝乳胰酶在溴氰菊酯抗性品系中表达量较高，且与蚊虫抗药性有关。侵害pallens。(c) 2005爱思唯尔公司版权所有。
Two serine proteinase genes were isolated from Culex pipiens pallens as significantly up-regulated genes in a deltamethrin-resistant strain through a combination of suppression substractive hybridization and gene expression profiling by macroarrays. These two genes were found to be expressed at least threefold higher in the resistant strain than in the susceptible one. By using rapid amplification of cDNA ends to screen the constructed cDNA library, we cloned these two sequences. There were 909 bp with an open reading frame of 786 bp in the sequence of trypsin cDNA (GenBank/NCBI AY034060), the deduced protein had 261 amino acids, which was most similar to the trypsin gene of Anopheles gambiae. There were 992 bp with an open reading frame of 816 bp in the chymotrypsin cDNA (GenBank/NCBI AF468495), and its deduced amino acid sequence had 271 amino acids, which was most similar to the chymotrypsin-like protein from Aedes aegypti. The two genes were stably expressed in mosquito C6/36 cells, and the expected 29 and 30 kDa bands were shown with Western blot, respectively. In these cells, after deltamethrin treatment, they had protective effects on the viability. The results indicate that trypsin and chymotrypsin were more highly expressed in the deltamethrin-resistant strain, and was related to insecticide resistance in mosquitoes, Cx. pipiens pallens. (c) 2005 Elsevier Inc. All rights reserved.