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茅苍术AlWRKY2基因的克隆及表达分析

植物生理学报 2024年第8期60卷 1253-1264页

作者：迟凯文管凤雅赵志强查良平安徽中医药大学药学院合肥230012 安徽省中医药科学院中药资源保护与开发研究所合肥230012

WRKY转录因子对植物生长过程中的物质代谢以及应对各类胁迫起着重要作用。本研究根据茅苍术转录组数据库,克隆得到WRKY2基因序列,命名为AlWRKY2(genBank登录号:OR343916)。利用软件对AlWRKY2基因进行生物信息学分析,显示该基因包含1个97... 详细信息

WRKY转录因子对植物生长过程中的物质代谢以及应对各类胁迫起着重要作用。本研究根据茅苍术转录组数据库,克隆得到WRKY2基因序列,命名为AlWRKY2(genBank登录号:OR343916)。利用软件对AlWRKY2基因进行生物信息学分析,显示该基因包含1个975 bp的开放阅读框,编码324个氨基酸,AlWRKY2蛋白理论等电点为6.62,理论分子质量为35426.90。蛋白质结构预测显示,AlWRKY2蛋白质二级结构主要由无规则卷曲构成。同源氨基酸序列分析表明茅苍术和软橡木树、杨梅、可可树、哥伦比亚锦葵等植物的WRKY2基因编码的氨基酸具有较高的同源性且都具有WRKY保守结构域。系统进化树分析表明茅苍术AlWRKY2氨基酸序列与菊科植物聚在同一分支,具有高度同源性;SDS-PAgE电泳结果显示AlWRKY2基因成功在大肠杆菌BL21(DE3)表达感受态中进行诱导表达,并在约50 kDa处表达出预期目的蛋白;含有AlWRKY2基因的重组菌在NaCl和甘露醇的胁迫条件下负调控菌落生长;利用qRT-PCR检测显示,在茅苍术中AlWRKY2的表达量在叶中最高,茎中最低,且在外源茉莉酸甲酯(MeJA)处理下提高了湖北茅苍术AlWRKY2基因表达量,并在24 h处表达量最高。亚细胞定位结果显示该基因位于细胞核中。本研究可为进一步研究茅苍术WRKY转录因子的功能提供参考。

关键词：茅苍术 WRKY转录因子基因克隆 qRT-PCR 表达分析亚细胞定位

茅苍术转录因子AlWRKY65的克隆与功能初探

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药学学报 2024年第5期59卷 1494-1502页

作者：管凤雅刘巍玮迟凯文曾凯玲谢晋查良平安徽中医药大学药学院安徽合肥230012 安徽医科大学药学院安徽合肥230032

WRKY转录因子是植物特有的一类转录因子,在植物多种生理过程中发挥重要的作用。本研究以茅苍术转录组数据为依据,对茅苍术AlWRKY基因与AlTPS基因的FPKM值进行相关性分析,结合分析结果筛选出与AlTPS1、AlTPS6基因表达量具有显著正相关性... 详细信息

WRKY转录因子是植物特有的一类转录因子,在植物多种生理过程中发挥重要的作用。本研究以茅苍术转录组数据为依据,对茅苍术AlWRKY基因与AlTPS基因的FPKM值进行相关性分析,结合分析结果筛选出与AlTPS1、AlTPS6基因表达量具有显著正相关性且FPKM值相对较高的候选基因AlWRKY65,对该候选基因进行克隆,获得AlWRKY65基因的开放阅读框(ORF),分析其编码蛋白的相关信息并进行基因表达研究。研究结果表明,AlWRKY65包含一个681bp的开放阅读框,编码226个氨基酸。AlWRKY65基因编码的氨基酸序列与向日葵HaWRKY65、莴苣LsWRKY65等植物基因编码的氨基酸序列具有较高同源性;该蛋白具有1个WRKYgQK保守结构域,属于WRKY基因家族的第IIe组;系统进化树分析显示,茅苍术AlWRKY65与洋蓟CcWRKY65蛋白亲缘关系较近;运用实时荧光定量PCR检测AlWRKY65基因在2个产地不同组织中的表达水平,结果表明,不同产地的茅苍术AlWRKY65基因具有组织表达差异性,均在叶片中表达量较高;在茉莉酸甲酯(MeJA)诱导处理48 h内,AlWRKY65基因的表达量下调;亚细胞定位和转录自激活活性分析显示,AlWRKY65定位于细胞核,不具有自激活活性。这些结果为进一步研究AlWRKY65在茅苍术中的生物学功能提供参考。

关键词：茅苍术 WRKY转录因子基因克隆表达分析亚细胞定位

基于优化的Maxent模型预测桔梗在中国的潜在适生区

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药学学报 2024年第9期59卷 2625-2633页

作者：张玉洁余函纹郑昭焕蒋超刘娟查良平池秀莲桂双英安徽中医药大学药学院安徽合肥230012 安徽中医药大学新安医学与中医药现代化研究所安徽合肥230012 中国中医科学院中药资源中心道地药材品质保障与资源持续利用全国重点实验室北京100700

桔梗Platycodongrandiflorum(Jacq.)***.作为最常用的大宗药材,在药用、食用和化妆品领域中都有着重要价值,市场需求量逐年增多,具有良好的开发前景。本研究以中国境内桔梗为研究对象,基于403条分布点和8个环境因子变量,运用参数优化后... 详细信息

桔梗Platycodongrandiflorum(Jacq.)***.作为最常用的大宗药材,在药用、食用和化妆品领域中都有着重要价值,市场需求量逐年增多,具有良好的开发前景。本研究以中国境内桔梗为研究对象,基于403条分布点和8个环境因子变量,运用参数优化后的Maxent模型探究未来气候条件下桔梗的适生分布情况。研究表明Maxent参数在特征组合(featurecombination,FC)为LQPH(L:线性特征;Q:二次型特征;P:片段化特征;H:乘积型特征),调控倍频(regularizationmultiplier,RM)为2.1时,模型模拟效果最佳,受试者曲线下面积(areaundercurve,AUC)为0.901;模型预测显示在当前气候条件下,桔梗广泛分布于中国的28个省份,适生区面积达2337419.98km^(2);在未来气候变化影响下,桔梗的适生区面积呈现降低趋势,分布中心整体呈现向北和向东偏移的趋势;当前三大主产区内桔梗的分布面积受气候变化影响的变化趋势不同,总体表现为在内蒙古赤峰的未来适生分布面积增加,而在共享社会经济路径5-8.5(shared socioeconomicpathways,SSP)排放情景下山东淄博和安徽亳州、太和县的未来适生分布面积减少甚至消失。建议维持和保护好适宜桔梗生长的保留区的生态环境和种质资源,增加在桔梗扩增区中的栽培研究,为后续拓展桔梗种植打下基础,由此促进桔梗资源的保护和可持续发展。

关键词：桔梗 Maxent模型参数优化气候变化适生区变化

Preparation and Identification of HLA-A*1101 Tetramer Loading with Human Cytomegalovirus pp65 Antigen Peptide

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Cellular & Molecular Immunology 2007年第2期4卷 141-146页

作者： Fengyao Li Lihui Xu Qingbing zha Xiaoyun chi Qiantao Jia Kianhui He Institute of Tissue Transplantation and Immunology College of Life Science and Technology Jinan University Guangzhou 510632 China Institute of Bioengineering College of Life Science and Technology Jinan University Guangzhou 510632 China Clinical Experimental Center of the First Affiliated Hospital Jinan University Guangzhou 510632 China

MHC/peptide tetramer technology has been widely used to study antigen-specific T cells, especially for identifying virus-specific CD8^＋ T cells in humans. The tetramer molecule is composed of HLA heavy chain, β2-microglobulin （β2m）, an antigenic peptide, and fluorescent-labeled streptavidin. To further investigate the HLA-A＊1101-restricted CD8^＋ T cell responses against human cytomegalovirus （HCMV）, we established an approach to prepare HLA-A＊1101 tetramer complexed with a peptide from HCMV. The cDNA encoding HLA-A＊1101 heavy chain was cloned and the prokaryotic expression vector for the ectodomain of HLA-A＊1101 fused with a BirA substrate peptide （HLA-A＊1101-BSP） at its carboxyl terminus was constructed. The fusion protein was highly expressed as inclusion bodies under optimized conditions in Escherichla coli. Moreover, HLA-A＊1101-BSP protein was refolded in the presence of β2m and an HCMV peptide pp6516.24 （gPISgHVLK, gPI）. Soluble HLA-A＊1101-gPI monomer was biotinylated and purified to a purity of 95%, which was subsequently combined with streptavidin to form tetramers at a yield of 〉 80%. The HLA-A＊1101-gPI tetramers could bind to virus-specific CD8^＋ T cells, suggesting soluble HLA-A＊1101-gPI tetramers were biologically functional. This study provides the basis for further evaluation of HLA-A＊1101-restricted CD8^＋ T cell responses against HCMV infection.

关键词： HLA-A* 1101 tetramer cytomegalovirus CD8^+T cell epitope

High Level Expression of HLA-A ~*0203-BSP Fusion Protein in Escherichia coli and Construction of Soluble HLA-A ~*0203 Monomer and Tetramer Loaded with Epstein-Barr Virus Peptide

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Cellular & Molecular Immunology 2007年第4期4卷 301-308页

作者： Qiantao Jia Lihui Xu Qingbing zha Xiaoyun chi Fengyao Li Xianhui He Institute of Tissue Transplantation and Immunology Institute of Bioengineering College of Life Science and Technology.Jinan University 601 Huangpu Dadao West Guangzhou 510632 China

Major histocompatibility complex （MHC） tetramer technology is critical for characterization of antigen-specific T cells. In the present study we reported the successful generation of HLA-A＊0203 tetramer loaded with Epstein- Barr virus EBNA3596-604 peptide （SVRDRLARL, SVR）. Prokaryotic expression vector for the ectodomain of the heavy chain of HLA-A＊0203 fused with a BirA substrate peptide （HLA-A＊0203-BSP） was constructed and the expression conditions of the fusion protein in Escherichia coli （E. coli） were optimized. The fusion protein was highly expressed in inclusion bodies within E. coil It was then refolded in the presence of 132-microglobulin and SVR peptide to form a soluble HLA-A＊0203-SVR monomer. After biotinylation with BirA, the monomer was purified by anion-exchange chromatography and its purity was up to 95%. The tetramer was then formulated by mixing the biotinylated monomer with streptavidin-PE at a ratio of 4：1. Flow cytometry showed that this tetramer could specifically react with antigen-specific CD8^＋ T cells, indicating that it was biologically functional. These results provide a foundation for further characterization of antigen-specific CD8^＋ T cells from HLA-A＊0203 subjects.

关键词： HLA-A*0203 tetramer Epstein-Barr virus prokaryotic expression biotinylation

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三相电压源型换流器故障耐受能力分析与评估

电力系统自动化 2016年第18期40卷 109-115页

作者：彭宇黄萌查晓明 chi K.TSE 武汉大学电气工程学院湖北省武汉市430072 香港理工大学电子与资讯工程系

三相电压源型并网换流器由于含有大量电力电子开关器件及相应控制环节,是一种强非线性系统。在交直流混合系统中受到电网侧故障或直流侧负载变化等扰动影响,大容量换流器将产生复杂的响应行为,给电网带来难以预计的影响。文中考虑电网... 详细信息

三相电压源型并网换流器由于含有大量电力电子开关器件及相应控制环节,是一种强非线性系统。在交直流混合系统中受到电网侧故障或直流侧负载变化等扰动影响,大容量换流器将产生复杂的响应行为,给电网带来难以预计的影响。文中考虑电网中三相电压暂降的典型故障,基于平均模型分析了三相电压源型换流器电流环的非线性特性。在此基础上,根据李雅普诺夫稳定性判据分析了故障前后换流器的稳定性,并以故障深度和故障持续时间为指标对换流器故障耐受能力进行评估。最后,通过并网系统的逐周期仿真验证了理论分析和计算的有效性。

关键词：并网换流器非线性分析李雅普诺夫稳定性故障耐受能力

Photoproduction of J/ψ in non-single-diffractive p+p collisions

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chinese Physics C 2019年第6期43卷 38-44页

作者： Ze-Hua Cao Li-Juan Ruan Ze-Bo Tang zha.g-Bu Xu chi Yang Shuai Yang Wang-Mei zha State Key Laboratory of Particle Detection and Electronics University of Science and Technology of China. Hefei 230026 China Department of Modem Physics University of Science and Technology of China Hefei 230026 China Brookhaven National Laboratory New York USA Shandong University Jinan 250100 China

Significant enhancements of J/ψ production at very low transverse momenta were recently observed by the ALICE and STAR collaborations in peripheral hadronic A+A collisions. The anomalous excess points to coherent photon-nucleus interactions in violent hadronic heavy-ion collisions, which were conventionally studied only in ultra-peripheral collisions. Assuming that the coherent photoproduction is the underlying mechanism responsible for the excess observed in peripheral A+A collisions, its contribution in p+p collisions with nuclear overlap, i.e. nonsingle-diffractive collisions, is of particular interest. In this paper, we perform a calculation of exclusive J/ψ photoproduction in non-single-diffractive p+p collisions at the RHIC and LHC energies based on the pQCD motivated parametrization using the world-wide experimental data, which could be further employed to improve the precision of the phenomenological calculations for photoproduction in A+A collisions. The differential rapidity and transverse momentum distributions of J/ψ from photoproduction are presented. In comparison with the J/ψ production from hadronic interactions, we find that the contribution of photoproduction is negligible.

关键词： photoproduction jpsi RHIC LHC

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基于剑桥模型的砂土剪胀模型

北京交通大学学报 2008年第1期32卷 49-53页

作者：迟明杰赵成刚北京交通大学土木建筑工程学院北京100044

对于中密砂和密砂,剪胀现象是一个比较显著的特征,相变线是描述剪胀现象的一条特征线,相变线比较容易确定.本文在已有弹塑性模型基础上,对硬化参数进行修正,并引入与相变线相关的状态参量,建立了一个新的剪胀性砂土本构模型.该模型尤其... 详细信息

对于中密砂和密砂,剪胀现象是一个比较显著的特征,相变线是描述剪胀现象的一条特征线,相变线比较容易确定.本文在已有弹塑性模型基础上,对硬化参数进行修正,并引入与相变线相关的状态参量,建立了一个新的剪胀性砂土本构模型.该模型尤其适合于中密砂和密砂,对于松砂则退化为原弹塑性模型.模型包含9个材料参数,参数比较容易确定.数值模拟结果能较好的描述砂土的主要特性.

关键词：岩土工程砂土本构模型硬化参数相变线状态参量

Dual ultra-wideband(UWB)radar-based sleep posture recognition system:Towards ubiquitous sleep monitoring

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再生工程(英文) 2023年第1期4卷 36-43页

作者： Derek Ka-Hei Lai Li-Wen zha Tommy Yau-Nam Leung Andy Yiu-Chau Tam Bryan Pak-Hei So Hyo-Jung Lim Daphne Sze Ki Cheung Duo Wai-chi Wong James Chung-Wai Cheung Department of Biomedical Engineering Faculty of EngineeringThe Hong Kong Polytechnic UniversityHong Kong 999077China Department of Bioengineering Imperial CollegeLondon SW72AZUnited Kingdom Research Institute of Smart Ageing The Hong Kong Polytechnic UniversityHong KongChina School of Nursing The Hong Kong Polytechnic UniversityHong KongChina

Sleep posture monitoring is an essential assessment for obstructive sleep apnea(OSA)*** objective of this study is to develop a machine learning-based sleep posture recognition system using a dual ultra-wideband radar *** collected radiofrequency data from two radars positioned over and at the side of the bed for 16 patients performing four sleep postures(supine,left and right lateral,and prone).We proposed and evaluated deep learning approaches that streamlined feature extraction and classification,and the traditional machine learning approaches that involved different combinations of feature extractors and *** results showed that the dual radar system performed better than either single *** statistical features with random forest classifier yielded the best accuracy(0.887),which could be further improved via an ablation study(0.938).Deep learning approach using transformer yielded accuracy of 0.713.

关键词： Obstructive sleep apnea Deep learning Sleep monitoring Feature extraction Ablation study