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Highly Efficient Base Editing in Viral Genome Based on Bacterial Artificial Chromosome Using a Cas9-Cytidine Deaminase Fused Protein

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Virologica Sinica 2020年第2期35卷 191-199页

作者： ke zheng Fang-Fang Jiang Le Su Xin Wang Yu-Xin Chen Huan-Chun Chen zheng-Fei Liu State key Laboratory of Agricultural Microbiology and key Laboratory of Preventive Veterinary Medicine in Hubei Province.College of Veterinary Medicine.Huazhong Agricultural University Wuhan 430070China Gene Editing Research Center Hebei University of Science and Technology.Shijiazhuang 050018China

Viruses evolve rapidly and continuously threaten animal health and economy,posing a great demand for rapid and efficient genome editing technologies to study virulence mechanism and develop effective *** present a highly efficient viral genome manipulation method using CRISPR-guided cytidine *** cloned pseudorabies virus genome into bacterial artificial chromosome,and used CRISPR-guided cytidine deaminase to directly convert cytidine(C)to uridine(U)to induce premature stop mutagenesis in viral *** editing efficiencies were 100%.Comprehensive bioinformatic analysis revealed that a large number of editable sites exist in pseudorabies virus(PRV)***,in our study viral genome exists as a plasmid in ***,suggesting that this method is virus *** application of base-editing provided an alternative approach to generate mutant virus and might accelerate study on virulence and vaccine development.

关键词： Pseudorabies virus(PRV) Bacterial artificial chromosome(BAC) Base-editing CRISPR7Cas9 Genome editing

Isolation and characterization of an isoamylase gene from rye

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The Crop Journal 2013年第2期1卷 127-133页

作者： ke zheng Jie Xu Qingtao Jiang André Laroche Yuming Wei Youliang zheng Zhenxiang Lu Lethbridge Research Centre Agriculture and Agri-Food Canada Triticeae Research Institute Sichuan Agricultural University

Genomic DNA and cDNA sequences of an isoamylase gene were isolated and characterized from the rye genome. The full-lengths of the rye isoamylase gene are 7351 bp for genomic DNA and 2364 bp for cDNA. There are 18 exons and 17 introns in the genomic sequence, which shares a similar organization with homologous genes from Aegilops tauschii, maize, rice and Arabidopsis. Exon regions of rye and other plant isoamylase genes are more conserved than the introns. High sequence similarity(>95%) was observed in mature proteins of isoamylase genes originating from rye, Ae. tauschii, wheat and barley. The transcript profile revealed that rye isoamylase is mainly expressed in the seed endosperm with a maximum level at the middle developmental stage(15 DPA). A phylogenetic tree based on the deduced aa sequences of mature proteins from rye and other plant isoamylases indicated that rye isoamylase is more closely related to Ae. tauschii wDBE1 and wheat iso1. This is the first report on identification and characterization of the isoamylase gene from rye, making it possible to explore the roles of this enzyme for amylopectin development in rye and triticale.

关键词： Secale cereale Isoamylase Gene structure Gene expression Phylogenetic tree

Phospho-proteomics identifies a critical role of ATF2 in pseudorabies virus replication

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Virologica Sinica 2022年第4期37卷 591-600页

作者： Fang-Fang Jiang Ren-Qi Wang Chao-Yue Guo ke zheng Hai-Long Liu Le Su Sheng-Song Xie Huan-Chun Chen zheng-Fei Liu State key Laboratory of Agricultural Microbiology Hongshan Laboratory and Key laboratory of Preventive Veterinary Medicine in Hubei ProvinceCollege of Veterinary MedicineHuazhong Agricultural UniversityWuhan430070China key Lab of Agricultural Animal Genetics Breedingand Reproduction of Ministry of EducationHuazhong Agricultural UniversityWuhan430070China

Pseudorabies virus(PRV),an etiological agent of pseudorabies in livestock,has negatively affected the porcine industry all over the *** cells are reported as the first site of PRV ***,the role of host proteins and its related signaling pathways in PRV replication is largely *** this study,we performed a quantitative phosphoproteomics screening on PRV-infected porcine kidney(PK-15)epithelial *** 5723phosphopeptides,corresponding to 2180 proteins,were obtained,and the phosphorylated states of 810 proteins were significantly different in PRV-infected cells compared with mock-infected cells(PkeGG analysis revealed that these differentially expressed phosphorylated proteins were predominantly related to RNA transport and MAPK signaling *** functional studies of NF-κB,transcription activator factor-2(ATF2),MAX and SOS genes in MAPK signaling pathway were analyzed using RNA interference(RNAi)*** showed that only ATF2-knockdown reduces both PRV titer and viral genome copy *** pathway inhibition and CRISPR/Cas9 gene knockout showed that ATF2 was required for the effective replication of PRV,especially during the biogenesis of viral genome ***,by overexpression of the ATF2 gene and point mutation of the amino acid positions 69/71 of ATF2,it was further demonstrated that the phosphorylation of ATF2 promoted PRV *** findings suggest that ATF2 may provide potential therapeutic target for inhibiting PRV infection.

关键词： Pseudorabies virus(PRV) Transcription activator factor-2(ATF2) iTRAQ Proteomics Phosphorylation

Chemical Chaperones Increasing Expression Level of Soluble Single-chain Fv Antibody(scFv2F3)

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Chemical Research in Chinese Universities 2007年第1期23卷 69-75页

作者： DU Xiu-bo SUN Ye LIN Feng zheng ke-yan WANG ke-wei LIN Ting-ting LIU Jun-qiu SHEN jia-cong LUO Gui-min key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education key Laboratory for Supramolecular Structure and Materials of the Ministry of Education Jilin University Changchun 130012 P. R. China

Glutathione peroxidase（GPX） is a sclenoenzyme that protects the biomembrane and other cellular components against oxidative damage. The selenium-containing single chain Fv fragment of monoelonal antibody 2173 （Se scFv2F3） is a kind of GPX mimic and it has a wide clinical applications because of its high activity and low antigenicity. Se-seFv2F3 is generated by the chemical modification of the single chain Fv fragment of monoelonal antibody 2F3 （ scFv2F3 ）, which can be expressed in E. coll. In this article, the effect of chemical chaperones, such as glycerol, glucose, and β-cyclodextrin added to the culture medium, on the expression of soluble scFv2F3 was investigated. The expression level was evaluated by the determination of soluble scFv2F3 contents in the whole cell lysates. The results suggest that both glycerol and β-cyclodextrin greatly increase the expression level of soluble scFv2F3, and β-cyclodextrin is found to be more effective compared with glycerol. Glucose has a slight effect on the expression level of soluble scFv2F3. This is the first example, wherein β-cyclodextrin has been used as a chemical chaperone during the cell culture to improve the expression level of recombinant proteins. In addition, chemical chaperones are found to decrease the toxic effect of IPTG on cells.

关键词： Glutathione peroxidase Soluble expression Glycerol Glucose β-Cyclodextrin

Effect of Polysaccharides from Fruit Body of Ganoderma tsugae on Bidirectional Regulation of Proinflammatory Cytokine Production in THP-1 Cells

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Chemical Research in Chinese Universities 2009年第4期25卷 487-491页

作者： LIU Ai-dong zheng ke-yan MIAO Qing-fang IkeJIMA Takashi ZHANG Jie FEI Xiao-fang College of Life Science Jilin University Changchun 130021 P. R. China College of Chemistry Jilin University Changchun 130021 P. R. China Department of Pharmacy the Affiliated Hospital Changchun University of Traditional Chinese Medicine Changchun 130021 P. R. China Department of Cardiology the Affiliated Hospital Changchun University of Traditional Chinese Medicine Changchun 130021 P. R. China China-Japan Research Institute of Medical and Pharmaceutical Sciences Shenyang Pharmaceutical University Shenyang 110016 R R. China

The polysaccharides,FIII-2,FIII-2-a,FIII-2-b and FIII-2-c were purified from the fruit body of Ganoderma tsugae（***） to study the effects of the four polysaccharides on the productions of human proinflammatory cytokines,interleukin-1α（IL-1α） and tumor necrosis factor α（TNFα）.The results show that the polysaccharides up-regulated the production of IL-1α and TNFα in THP-1 cells under non-stimulatory condition or under stimulation of both lower concentration lipopolysaccharide（LPS） and phorbol myristate acetate（PMA）,and they down-regulated the secretion of IL-1α and TNFα under stronger stimulatory *** four polysaccharides promoted the expression of TNFα *** of TNFα protein reflects the quantity of TNFα *** polysaccharides purified from Ganoderma tsugae bodies exhibit bidirectional regulation effects on cytokine production under different cell stimulatory conditions.

关键词： Polysaccharide Bidirectional regulation Cytokine

A new flavonol glycoside from Hydrangea macrophylla (Thunb.) Seringe

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Chinese Chemical Letters 2010年第6期21卷 690-692页

作者： Wei Sheng Feng Yan Li Zhang Xiao ke zheng Bo Zhu Li Gong Yang Zhen Li Henan College of Traditional Chinese Medicine Zhengzhou 450008China

A new kaempferol glycoside,named kaempferol 3-O-[6″-O-β-D-glucopyranosyl-6′′′-O-α-L-rhamnopy ranosyl-2′′′′-O-α-L-rhamnopyranosyl]-β-D-glucopyranoside,was isolated from Hydrangea macrophylla(Thunb.) *** str... 详细信息

关键词： Hydrangea macrophylla (Thunb.) Seringe Kaempferol glycoside

Tin-induced microstructural changes and associated corrosion resistance of biodegradable Mg–Zn alloy

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Rare Metals 2022年第3期41卷 883-888页

作者： Yun Gao Ping Zhao Xiang-Qian Cao R.Devesh K.Misra Wei Wang ke-zheng Chen Laboratory of Functional and Biomedical Nanomaterials College of Materials Science and EngineeringQingdao University of Science and TechnologyQingdao 266042China Shandong Provincial University Laboratory for Protected Horticulture&Architecture and Art College Weifang University of Science and TechnologyWeifang 262700China Department of Metallurgical Materials and Biomedical EngineeringUniversity of TexasEl PasoTX 79968USA

As a new degradable medical metal material, Mg–Zn alloy has been widely used in clinical application. However, its popularization and application are seriously restricted by poor corrosion performance in the biological liquid environment. For this reason, we added a relatively non-toxic element to the human body (Sn) to Mg–Zn alloy, and we prepared Mg–2Zn–3Sn ternary system alloy.

关键词： alloy corrosion alloy.

Particle size distribution of coal and gangue after impact-crush separation

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Journal of Central South University 2017年第6期24卷 1252-1262页

作者： YANG Dao-long LI Jian-ping DU Chang-long zheng ke-hong LIU Song-yong School of Mechatronic Engineering China University of Mining & Technology College of Mines and Earth Sciences University of Utah

Based on the separation and backfilling system of coal and gangue, the mineral material impact experiments were conducted utilizing the hardness difference between coal and gangue according to the uniaxial compression experiments. The broken coal and gangue particles were collected and screened by different size meshes. The particle size distributions of coal and gangue under different impact velocities were researched according to the Rosin-Rammler distribution. The relationships between separation indicators and impact velocities were discussed. It is found from experiments that there is a fully broken boundary of coal material. The experimental results indicate that the Rosin-Rammler distribution could accurately describe the particle size distribution of broken coal and gangue under different impact velocities, and there is a minimum overlap region when the impact velocity is 12.10 m/s which leads to the minimum mixed degree of coal and gangue, and consequently the benefit of coal and gangue separation.

关键词： Rosin-Rammler distribution impact crush separation indicator coal and gangue separation and backfilling system

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山区复杂地形下路基或桥梁方案判别模型研究

湖南大学学报（自然科学版） 2009年第3期36卷 22-25页

作者：郑柯郑勇湖南大学土木工程学院湖南长沙410082

针对山区复杂地形下高速公路布线的多方案选择问题,拟定数个能贴切描述地形纵、横向起伏变化的指标,以已有的优秀山区高速公路路线方案为依托采集样本数据,并对其进行判别分析,最终建立并标定了山区高速公路路线设计中路基和桥梁方案的... 详细信息

针对山区复杂地形下高速公路布线的多方案选择问题,拟定数个能贴切描述地形纵、横向起伏变化的指标,以已有的优秀山区高速公路路线方案为依托采集样本数据,并对其进行判别分析,最终建立并标定了山区高速公路路线设计中路基和桥梁方案的判别模型.通过把模型应用到其他已设计的优秀山区高速公路路线方案上,得到与实际方案相一致的判别结果.

关键词：山区高速公路数字地形路基桥梁判别分析