Serine proteinase over-expression in relation to deltamethrin resistance in Culex pipiens pallens

Gong, Maoqing; Shen, Bo; Gu, Yan; Tian, Haihua; Ma, Lei; Li, Xiulan; Yang, Mingxia; Hao, Ying; Sun, Yan; Hu, Xiaobang; Li, Jianyong; Zhu, Changliang

doi:10.1016/j.abb.2005.03.021

Cited by 49 publications

(45 citation statements)

References 22 publications

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“…Trypsin and alpha-chymotrypsin, which shared similar properties of the insect midgut alkaline proteases, rapidly degraded the polh (polyhedrin) protein in vitro, leaving only the toxic Cry1Ac protein behind (Seo et al 2005). We also observed that NYD-Tr and NYD-Ch were associated with deltemathrin resistance of C. pipiens pallens (Gong et al 2005). After the expression and purification of NYD-Tr and NYD-Ch, we got the actual molecular weight of NYD-Tr and NYD-Ch, which were similar to trypsins and chymo- trypsins from other insects and mammals (Lehane and Billingsley 1996;Barrett et al 1998).…”

Section: Discussionsupporting

confidence: 52%

“…Using rapid amplification of cDNA ends (RACE) from the constructed cDNA library, we obtained full-length cDNA of these two sequences (GenBank/NCBI AY034060, GenBank/NCBI AF468495). Stable transfection of these two genes into mosquito C6/36 cells conferred protective effects (Gong et al 2005), while RNAi-mediated knockdown made C6/36 cells more sensitive to deltamethrin (Gong et al, unpublished). These results suggested that trypsin-and chymotrypsin-like proteases might be directly involved in the metabolism of deltamethrin.…”

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confidence: 99%

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Expression and characterization of two pesticide resistance-associated serine protease genes (NYD-tr and NYD-ch) from Culex pipiens pallens for metabolism of deltamethrin

et al. 2008

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Two deltamethrin resistance-associated serine protease genes (NYD-tr and NYD-ch) were isolated from Culex pipiens pallens in our previous study. To study the function of NYD-Tr and NYD-Ch in the metabolism of deltamethrin, we constructed the recombinant plasmid pET32a(+)/NYD-tr and pET32a(+)/NYD-ch with a 6x histidine tag. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analyses of the recombinant proteins revealed that the molecular weights of NYD-Tr and NYD-Ch are 42 and 50 kDa. Enzyme activity assay indicated that the recombinant NYD-Tr and NYD-Ch had the corresponding features of trypsin and chymotrypsin. Using BApNA as the substrate, NYD-Tr gave optimal activity between pH 9.0 and 10.5, while NYD-Ch was optimally active over the range of pH 8.0-11.0 using the S(Ala)(2)ProPhe-pNA as the substrate. Then, we investigated the metabolism of deltamethrin by NYD-Tr and NYD-Ch. Our results showed that NYD-Tr and NYD-Ch could hydrolyze deltamethrin. The acute oral toxicity of the metabolite to Wistar rats was much lower than deltamethrin.

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Section: Discussionsupporting

confidence: 52%

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confidence: 99%

Expression and characterization of two pesticide resistance-associated serine protease genes (NYD-tr and NYD-ch) from Culex pipiens pallens for metabolism of deltamethrin

et al. 2008

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“…So there is a possibility that the expression of NYD-OP7 at the protein level is not as high as the mRNA level which may be the reason for why high NYD-OP7 transcriptional level expression only causes a modest increase in resistance. In a recent study, we have proved that the cotransfection of two genes (NYD-Tr and NYD-Ch) into C6/36 cells can induce deltamethrin resistance [33]. All these data suggest that insecticide resistance is more complex than we considered and we believe that further investigation should be done to fully elucidate the role that NYD-OP7 has in insecticide resistance.…”

Section: Discussionmentioning

confidence: 88%

Cloning and characterization of NYD-OP7, a novel deltamethrin resistance associated gene from Culex pipiens pallens

Sun

Wang

et al. 2007

Pesticide Biochemistry and Physiology

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One mosquito opsin gene, NYD-OP7, has been cloned from Culex pipiens pallens. An open reading frame (ORF) of 1116 bp was found to encode a putative 371 amino acids protein which exhibits high identity with opsins from Aedes and Anopheles mosquitoes. Transcript expression of NYD-OP7 was determined by real-time PCR in all life stages of deltamethrin-susceptible and -resistant strains of the Culex mosquito. The results demonstrated that this gene is expressed at all developmental stages, and it is expressed predominantly at the pupae and adult stages. Meanwhile, in pupae and adults, NYD-OP7 is overexpressed in deltamethrin-resistant strain than in -susceptible strain. Importantly, stable expression of NYD-OP7 in the mosquito C6/36 cells can confer moderate deltamethrin resistantce. Our study provides the Wrst direct evidence that increased expression of an opsin gene may play some role in the development of deltamethrin resistantce in Cx. pipiens pallens.

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“…As Gong et al reported in 2005, the deltamethrin resistance of mosquito cell line was up-regulated by cotransfection of trypsin and chymotrysin gene. However, the changes were not observed with separate transfection (Gong et al 2005). In our experiment, we up-regulated GBE gene in C6/36 cells alone, thus may not lead to higher resistance to deltamethrin without cooperation of other genes.…”

Section: Discussionmentioning

confidence: 65%

Glycogen branching enzyme: a novel deltamethrin resistance-associated gene from Culex pipiens pallens

Yang

Sun

et al. 2008

Parasitol Res

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The glycogen branching enzyme (GBE) gene has been cloned from Culex pipiens pallens. An open reading frame of 2,070 bp was found to encode a putative 689 amino acid protein. The deduced amino acid sequence shares 82% and 60% identity with GBE genes from Aedes aegypti and Homo sapiens. Transcript expression of GBE was determined by real-time polymerase chain reaction in deltamethrin-susceptible and deltamethrin-resistant strains of the Culex mosquito. The results demonstrated that this gene has 19.7-fold higher expression in deltamethrin-resistant strain. Unexpectedly, the following study showed that cell proliferation decreased obviously in the GBE overexpression group compared to the empty vector control or the unrelated gene when treated by deltamethrin in the mosquito C6/36 cells whose mechanism is still unexplained. Our study provides the first direct evidence that GBE gene may play some role in the development of deltamethrin resistance in C. pipiens pallens.

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Serine proteinase over-expression in relation to deltamethrin resistance in Culex pipiens pallens

Cited by 49 publications

References 22 publications

Expression and characterization of two pesticide resistance-associated serine protease genes (NYD-tr and NYD-ch) from Culex pipiens pallens for metabolism of deltamethrin

Expression and characterization of two pesticide resistance-associated serine protease genes (NYD-tr and NYD-ch) from Culex pipiens pallens for metabolism of deltamethrin

Cloning and characterization of NYD-OP7, a novel deltamethrin resistance associated gene from Culex pipiens pallens

Glycogen branching enzyme: a novel deltamethrin resistance-associated gene from Culex pipiens pallens

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