Wen-Jing Zhao, Yan Li, Zhen-Long Jiao, Pei-Pei Su, Long-Bing Yang, Chao-Qin Sun, Jiang-Fan Xiu, Xiao-Li Shang, Guo Guo
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The results showed that <i>MdCht9</i> consists of three domains, highly expressed in a larval salivary gland. RNAi silencing of <i>MdCht9</i> resulted in significant down-regulation of chitin content and expression of 15 chitin-binding protein (CBP) genes, implying a new insight that <i>MdCht9</i> might regulate chitin content by influencing the expression of CBPs. In addition, more than half of the lethality and partial wing deformity appeared due to the <i>dsMdCht9</i> treatment. In addition, the rMdCht9 exhibited anti-microbial activity towards <i>Candida albicans</i> (fungus) but not towards <i>Escherichia coli</i> (G−) or <i>Staphylococcus aureus</i> (G+). 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引用次数: 0
摘要
昆虫几丁质酶被认为是害虫控制的潜在目标。在这项工作中,研究人员发现来自家蝇的一种新型第 IV 组几丁质酶基因 MdCht9 在生理活动中具有多种功能,包括几丁质调节、发育和抗真菌免疫。对 MdCht9 基因进行了克隆和测序,分析了其系统发育,并测定了其在正常幼虫和 20E 处理幼虫中的表达。随后,进行了 RNA 干扰(RNAi)介导的 MdCht9 基因敲除,并进行了生化检测、形态观察和转录组分析。最后,纯化重组蛋白 MdCht9(rMdCht9),并检测其抗微生物活性和酶特性。结果表明,MdCht9由三个结构域组成,在幼虫唾液腺中高度表达。RNAi沉默MdCht9可显著下调几丁质含量和15个几丁质结合蛋白(CBP)基因的表达,这意味着MdCht9可能通过影响CBP的表达来调控几丁质含量。此外,一半以上的致死和翅膀部分畸形是由于dsMdCht9处理造成的。此外,rMdCht9 对白色念珠菌(真菌)具有抗微生物活性,但对大肠杆菌(G-)或金黄色葡萄球菌(G+)没有抗微生物活性。我们的工作拓展了以往对几丁质酶的研究,同时为害虫管理提供了一个潜在的目标。
Function analysis and characterisation of a novel chitinase, MdCht9, in Musca domestica
Insect chitinases have been proposed as potential targets for pest control. In this work, a novel group IV chitinase gene, MdCht9, from Musca domestica was found to have multiple functions in the physiological activity, including chitin regulation, development and antifungal immunity. The MdCht9 gene was cloned and sequenced, its phylogeny was analysed and its expression was determined in normal and 20E treated larvae. Subsequently, RNA interference (RNAi)-mediated MdCht9 knockdown was performed, followed by biochemical assays, morphological observations and transcriptome analysis. Finally, the recombinant protein MdCht9 (rMdCht9) was purified and tested for anti-microbial activity and enzyme characteristics. The results showed that MdCht9 consists of three domains, highly expressed in a larval salivary gland. RNAi silencing of MdCht9 resulted in significant down-regulation of chitin content and expression of 15 chitin-binding protein (CBP) genes, implying a new insight that MdCht9 might regulate chitin content by influencing the expression of CBPs. In addition, more than half of the lethality and partial wing deformity appeared due to the dsMdCht9 treatment. In addition, the rMdCht9 exhibited anti-microbial activity towards Candida albicans (fungus) but not towards Escherichia coli (G−) or Staphylococcus aureus (G+). Our work expands on previous studies of chitinase while providing a potential target for pest management.
期刊介绍:
Insect Molecular Biology has been dedicated to providing researchers with the opportunity to publish high quality original research on topics broadly related to insect molecular biology since 1992. IMB is particularly interested in publishing research in insect genomics/genes and proteomics/proteins.
This includes research related to:
• insect gene structure
• control of gene expression
• localisation and function/activity of proteins
• interactions of proteins and ligands/substrates
• effect of mutations on gene/protein function
• evolution of insect genes/genomes, especially where principles relevant to insects in general are established
• molecular population genetics where data are used to identify genes (or regions of genomes) involved in specific adaptations
• gene mapping using molecular tools
• molecular interactions of insects with microorganisms including Wolbachia, symbionts and viruses or other pathogens transmitted by insects
Papers can include large data sets e.g.from micro-array or proteomic experiments or analyses of genome sequences done in silico (subject to the data being placed in the context of hypothesis testing).