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| 1 | Proteomics analysis of Trichoplusia ni midgut epithelial cell brush border membrane vesicles显示文摘The insect midgut epithelium is composed of columnar, goblet, and regenerative cells. Columnar epithelial cells are the most abundant and have membrane protrusions that form the brush border membrane (BBM) on their apical side. These increase surface area available for the transport of nutrients, but also provide opportunities for interaction with xenobiotics such as pathogens, toxins and host plant allelochemicals. Recent improvements in proteomic and bioinfbrmatics tools provided an opportunity to determine the proteome of the T. ni BBM in unprecedented detail. This study reports the identification of proteins from BBM vesicles (BBMVs) using single dimension polyacrylamide gel elec? trophoresis coupled with multi-dimensional protein identification technology. More than 3000 proteins were associated with the BBMV of which 697 were predicted to possess either a signal peptide, at least one transmembrane domain or a GPI-anchor signal. Of these, bioinfbrmatics analysis and manual curation predicted that 185 may be associated with the BBMV or epithelial cell plasma membrane. These are discussed with respect to their predicted functions, namely digestion, nutrient uptake, cell signaling, development, cell-cell interactions, and other functions. We believe this to be the most detailed proteomic analysis of the lepidopteran midgut epithelium membrane to date, which will provide information to better understand the biochemical, physiological and pathological processes taking place in the larval midgut. | Muhammad Afzal Javed Cathy Coutu David A. Theilmann Martin A. Erlandson Dwayne D. Hegedus | 2019 | Insect Science2019，26，3： | 1 |
| 2 | siRNA抑制PTTG1表达对人脑胶质瘤细胞增殖、侵袭性的影响显示文摘目的构建针对人脑胶质瘤细胞系的高效沉默PTTG1的RNAi载体;探讨PTTG1 siRNA干扰质粒对人脑胶质瘤细胞系U373增殖、侵袭性的影响。方法设计三对PTTG1基因干扰序列,合成能够特异干扰PTTG1的siRNA,然后将其与p Genesil2载体连接,构建pGenesil2-PTTG1 siRNA干扰载体,在脂质体作用下将PTTG1 siRNA质粒转染至胶质瘤U373细胞后,用RT-PCR和Western blot方法检测PTTG1 mRNA及蛋白表达水平的变化,通过细胞增殖实验、划痕实验和Transwell小室实验检测PTTG1对胶质瘤细胞增殖、迁移及侵袭能力的影响。结果在胶质瘤U373细胞转染3个不同的siRNA片段干扰PTTG1的表达,RTPCR检测结果显示,PTTG1 siRNA1干扰质粒对PTTG1表达的抑制作用最明显[(0.47±0.12)vs(1.00±0.15),P<0.01]。转染PTTG1 siRNA质粒的胶质瘤细胞U373在48h和72h细胞增殖能力均显著低于对照组。细胞划痕实验显示干扰PTTG1可明显缩短U373细胞的迁移距离;Transwell小室结果显示干扰PTTG1后U373的穿膜细胞百分率明显下降[(58.00±8.72)%vs(36.3±7.76)%,P<0.05]。结论 PTTG1 siRNA干扰质粒可明显抑制胶质瘤U373细胞PTTG1的表达,并可抑制胶质瘤细胞的增殖、侵袭及迁移能力。 | 闫海成 窦长武 田复明 妥少勇 王薇 | 2015 | 中国医药科学2015，5，21： | 0 |
| 3 | Integrated transcriptome sequencing and RNA interference reveals molecular changes in Diaphorina citri after exposure to validamycin显示文摘Validamycin has been widely used as a specific competitive inhibitor of tre-halase.In our previous research,validamycin significantly inhibited trehalase activity and chitin synthesis in Diaphorina citri,resulting in abnormal phenotypes.However,the mechanism of validamycin's action on D.citri remains unclear.Here,using a comparative transcriptome analysis,464 differentially expressed genes(DEGs)in D.citri were identified after validamycin treatment.A Gene Ontology enrichment analysis revealed that these DEGs were mainly involved in“small molecule process”,“structural molecule ac-tivity”and“transition metal ion binding”.DEGs involved in chitin metabolism,cuticle synthesis and insecticide detoxification were validated by reverse transcription quantitative polymerase chain reaction.The RNA interference of D.citri chitinase-like protein EN03 and D.citri cuticle protein 7 genes significantly affected D.citri molting.Moreover,the recombinant chitinase-like protein EN03 exhibited a chitin-binding property,and an antimicrobial activity against Bacillus subtilis.This study provides a first insight into the molecular changes in D.citri after exposure to validamycin and identifies two effective RNA interference targets for D.citri control. | Hai-Zhong Yu Yan-Xin Xie Jie Wang Ying Wang Yi-Min Du He-Gui Wang Ba-Lian Zhong Bo Zhu Xiu-Dao Yu Zhan-Jun Lu | 2021 | Insect Science2021，28，6： | 0 |
| 4 | The coming of RNA-based pest controls显示文摘RNA interference( RNAi) is a gene silencing tool that targets messenger RNA( m RNA)transcripts in a sequence specific manner and down regulates gene expression by interacting with small interfering RNAs( si RNAs). m RNAs can be silenced either through endogenous degradation via nuclease activity( e. g.,RNAi pathway),or by inhibiting translation( e. g.,mi RNA). Over the past decade,RNAi has been used broadly in entomological research to decipher the gene functions in insects. With the success of RNAi in functional genomics research,much attention has shifted to the potential applications of RNAi in agriculture,especially for the control of insect pests. In addition to RNAi-based gene silencing,genome editing,an exciting new biotechnology,offers yet another option for pest controls. The modification of plant genomes to knock-in genes that are heritable to increase the tolerance of plants to insect infestation,or knock-out genes in pest insects through genetically modified( GM)-insect releases are on the forefront of the genetic-based pest managements. In this review,we summarize the current knowledge regarding RNAi-based gene silencing and CRISPR / Cas9-based genome editing. Technical challenges and regulatory concerns for this new wave of RNA-based pest controls are discussed in great detail. We also share the perspective of modifying current environmental risk assessment frameworks to better fit the RNA-based pest management strategies. Given the current discussions / attentions over the safety of Genetically Modified Organisms( GMO),we,respectively,compared pros and cons of RNAibased gene silencing and CRISPR / Gas9-based genome editing,and later we identified the regulatory issues that should be addressed before these emerging biotechnologies can move from the bench top to the table top. | Xu Linghua Zeng Baosheng Noland Jeffery E. Huang Yongping Zhou Xuguo | 2015 | 植物保护学报2015，42，5： | 0 |
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