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Research on Multi-modal In-Vehicle Intelligent Personal Assistant Design
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作者 WANG Jia-rou TANG Cheng-xin SHUAI Liang-ying 《印刷与数字媒体技术研究》 CAS 北大核心 2024年第4期136-146,共11页
Intelligent personal assistants play a pivotal role in in-vehicle systems,significantly enhancing life efficiency,driving safety,and decision-making support.In this study,the multi-modal design elements of intelligent... Intelligent personal assistants play a pivotal role in in-vehicle systems,significantly enhancing life efficiency,driving safety,and decision-making support.In this study,the multi-modal design elements of intelligent personal assistants within the context of visual,auditory,and somatosensory interactions with drivers were discussed.Their impact on the driver’s psychological state through various modes such as visual imagery,voice interaction,and gesture interaction were explored.The study also introduced innovative designs for in-vehicle intelligent personal assistants,incorporating design principles such as driver-centricity,prioritizing passenger safety,and utilizing timely feedback as a criterion.Additionally,the study employed design methods like driver behavior research and driving situation analysis to enhance the emotional connection between drivers and their vehicles,ultimately improving driver satisfaction and trust. 展开更多
关键词 Intelligent personal assistants Multi-modal design User psychology in-vehicle interaction Voice interaction Emotional design
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Phosphodiesterase 9 localization in cytoplasm and nucleus:the gateway to selective targeting in neuroprotection?
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作者 Giovanni Ribaudo Matteo Giannangeli +1 位作者 Margrate Anyanwu Alessandra Gianoncelli 《Neural Regeneration Research》 SCIE CAS 2025年第6期1701-1702,共2页
The umbrella term"neurodege ne rative disorders"(NDDs) refers to several conditions characterized by a progressive loss of structure and function of cells belonging to the nervous system.Such diseases affect... The umbrella term"neurodege ne rative disorders"(NDDs) refers to several conditions characterized by a progressive loss of structure and function of cells belonging to the nervous system.Such diseases affect more than 50million people worldwide.Neurodegenerative disorders are characterized by sundry factors and pathophysiological mechanisms that a re challenging to be fully profiled.Many of these rely on cell signaling pathways to preserve homeostasis,involving second messengers such as cyclic adenosine monophosphate (cAMP)and cyclic guanosine 3',5'-monophosphate(cGMP).Their ability to control the duration and amplitude of the signaling cascade is given by the presence of several common and uncommon effectors.Protein kinases A and G (PKA and PKG),phosphodiesterases (PDEs),and scaffold proteins are among them. 展开更多
关键词 gateway HOMEOSTASIS preserve
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Measurement of in-vehicle volatile organic compounds under static conditions 被引量:28
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作者 YOU Ke-wei GE Yun-shan +4 位作者 HU Bin NING Zhan-wu ZHAO Shou-tang ZHANG Yan-ni XIE Peng 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2007年第10期1208-1213,共6页
The types and quantities of volatile organic compounds (VOCs) inside vehicles have been determined in one new vehicle and two old vehicles under static conditions using the Thermodesorber-Gas Chromatograph/Mass Spec... The types and quantities of volatile organic compounds (VOCs) inside vehicles have been determined in one new vehicle and two old vehicles under static conditions using the Thermodesorber-Gas Chromatograph/Mass Spectrometer (TD-GC/MS). Air sampling and analysis was conducted under the requirement of USEPA Method TO-17. A room-size, environment test chamber was utilized to provide stable and accurate control of the required environmental conditions (temperature, humidity, horizontal and vertical airflow velocity, and background VOCs concentration). Static vehicle testing demonstrated that although the amount of total volatile organic compounds (TVOC) detected within each vehicle was relatively distinct (4940 μg/m^3 in the new vehicle A, 1240 μg/m^3 in used vehicle B, and 132 μg/m^3 in used vehicle C), toluene, xylene, some aromatic compounds, and various C7-C12 alkanes were among the predominant VOC species in all three vehicles tested. In addition, tetramethyl succinonitrile, possibly derived from foam cushions was detected in vehicle B. The types and quantities of VOCs varied considerably according to various kinds of factors, such as, vehicle age, vehicle model, temperature, air exchange rate, and environment airflow velocity. For example, if the airflow velocity increases from 0.1 m/s to 0.7 m/s, the vehicle's air exchange rate increases from 0.15 h^-1 to 0.67 h^-1, and in-vehicle TVOC concentration decreases from 1780 to 1201 μg/m^3. 展开更多
关键词 volatile organic compounds (VOCs) in-vehicle trim material static conditions
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A novel intrusion detection model for the CAN bus packet of in-vehicle network based on attention mechanism and autoencoder 被引量:2
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作者 Pengcheng Wei Bo Wang +2 位作者 Xiaojun Dai Li Li Fangcheng He 《Digital Communications and Networks》 SCIE CSCD 2023年第1期14-21,共8页
The attacks on in-vehicle Controller Area Network(CAN)bus messages severely disrupt normal communication between vehicles.Therefore,researches on intrusion detection models for CAN have positive business value for veh... The attacks on in-vehicle Controller Area Network(CAN)bus messages severely disrupt normal communication between vehicles.Therefore,researches on intrusion detection models for CAN have positive business value for vehicle security,and the intrusion detection technology for CAN bus messages can effectively protect the invehicle network from unlawful attacks.Previous machine learning-based models are unable to effectively identify intrusive abnormal messages due to their inherent shortcomings.Hence,to address the shortcomings of the previous machine learning-based intrusion detection technique,we propose a novel method using Attention Mechanism and AutoEncoder for Intrusion Detection(AMAEID).The AMAEID model first converts the raw hexadecimal message data into binary format to obtain better input.Then the AMAEID model encodes and decodes the binary message data using a multi-layer denoising autoencoder model to obtain a hidden feature representation that can represent the potential features behind the message data at a deeper level.Finally,the AMAEID model uses the attention mechanism and the fully connected layer network to infer whether the message is an abnormal message or not.The experimental results with three evaluation metrics on a real in-vehicle CAN bus message dataset outperform some traditional machine learning algorithms,demonstrating the effectiveness of the AMAEID model. 展开更多
关键词 Controller area network bus packet in-vehicle network Attention mechanism Autoencoder
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Recent Trends of In-Vehicle Time Sensitive Networking Technologies, Applications and Challenges 被引量:2
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作者 Yanli Xu Jian Shang Hao Tang 《China Communications》 SCIE CSCD 2023年第11期30-55,共26页
With the vigorous development of automobile industry,in-vehicle network is also constantly upgraded to meet data transmission requirements of emerging applications.The main transmission requirements are low latency an... With the vigorous development of automobile industry,in-vehicle network is also constantly upgraded to meet data transmission requirements of emerging applications.The main transmission requirements are low latency and certainty especially for autonomous driving.Time sensitive networking(TSN)based on Ethernet gives a possible solution to these requirements.Previous surveys usually investigated TSN from a general perspective,which referred to TSN of various application fields.In this paper,we focus on the application of TSN to the in-vehicle networks.For in-vehicle networks,we discuss all related TSN standards specified by IEEE 802.1 work group up to now.We further overview and analyze recent literature on various aspects of TSN for automotive applications,including synchronization,resource reservation,scheduling,certainty,software and hardware.Application scenarios of TSN for in-vehicle networks are analyzed one by one.Since TSN of in-vehicle network is still at a very initial stage,this paper also gives insights on open issues,future research directions and possible solutions. 展开更多
关键词 automobile industry deterministic transmission in-vehicle network low latency time sensitive networking(TSN)
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Hybrid In-Vehicle Background Noise Reduction for Robust Speech Recognition:The Possibilities of Next Generation 5G Data Networks
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作者 Radek Martinek Jan Baros +2 位作者 Rene Jaros Lukas Danys Jan Nedoma 《Computers, Materials & Continua》 SCIE EI 2022年第6期4659-4676,共18页
This pilot study focuses on employment of hybrid LMS-ICA system for in-vehicle background noise reduction.Modern vehicles are nowadays increasingly supporting voice commands,which are one of the pillars of autonomous ... This pilot study focuses on employment of hybrid LMS-ICA system for in-vehicle background noise reduction.Modern vehicles are nowadays increasingly supporting voice commands,which are one of the pillars of autonomous and SMART vehicles.Robust speaker recognition for context-aware in-vehicle applications is limited to a certain extent by in-vehicle back-ground noise.This article presents the new concept of a hybrid system which is implemented as a virtual instrument.The highly modular concept of the virtual car used in combination with real recordings of various driving scenarios enables effective testing of the investigated methods of in-vehicle background noise reduction.The study also presents a unique concept of an adaptive system using intelligent clusters of distributed next generation 5G data networks,which allows the exchange of interference information and/or optimal hybrid algorithm settings between individual vehicles.On average,the unfiltered voice commands were successfully recognized in 29.34%of all scenarios,while the LMS reached up to 71.81%,and LMS-ICA hybrid improved the performance further to 73.03%. 展开更多
关键词 5G noise reduction hybrid algorithms speech recognition 5G data networks in-vehicle background noise
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Chip Design for In-Vehicle System Transmitter
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作者 Majeed Nader John Liu 《Journal of Computer and Communications》 2018年第5期66-82,共17页
This paper presents embedded system design of the In-Vehicle System (IVS) for the European Union (EU) emergency call (eCall) system. The IVS transmitter modules are designed, developed and implemented on a field progr... This paper presents embedded system design of the In-Vehicle System (IVS) for the European Union (EU) emergency call (eCall) system. The IVS transmitter modules are designed, developed and implemented on a field programmable gate array (FPGA) device. The modules are simulated, synthesized, and optimized to be loaded on a reconfigurable device as a system-on-chip (SoC) for the IVS electronic device. All the modules of the transmitter are designed as a single embedded module. The bench-top test is completed for testing and verification of the developed modules. The hardware architecture and interfaces are discussed. The IVS signal processing time is analyzed for multiple frequencies. A range of appropriate frequency and two hardware interfaces are proposed. A state-of-the-art FPGA design is employed as a first implementation approach for the IVS prototyping platform. This work is used as an initial step to implement all the modules of the IVS on a single SoC chip. 展开更多
关键词 EU Emergency-Call (eCall) FPGA in-vehicle System TRANSMITTER SYSTEM-ON-CHIP
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An Energy Efficient Routing Protocol for In-Vehicle Wireless Sensor Networks
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作者 Chundong Wang Zhentang Zhao +1 位作者 Likun Zhu Honglei Yao 《国际计算机前沿大会会议论文集》 2017年第2期39-40,共2页
In this paper, an advanced distributed energy-efficient clustering (ADEEC) protocol was proposed with the aim of balancing energy consumption across the nodes to achieve longer network lifetime for In-Vehicle Wireless... In this paper, an advanced distributed energy-efficient clustering (ADEEC) protocol was proposed with the aim of balancing energy consumption across the nodes to achieve longer network lifetime for In-Vehicle Wireless Sensor Networks (IVWSNs). The algorithm changes the cluster head selection probability based on residual energy and location distribution of nodes. Then node associate with the cluster head with least communication cost and high residual energy. Simulation results show that ADEEC achieves longer stability period, network lifetime,and throughput than the other classical clustering algorithms. 展开更多
关键词 in-vehicle WIRELESS Sensor Network ROUTING PROTOCOL CLUSTERING Energy EFFICIENCY
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Ancient Zhenyuan, Gateway to East Guizhou
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作者 MENG JIAXIN 《China Today》 2024年第1期76-80,共5页
This ancient city has a history going back over 2,000 years.It has witnessed the fusion of multiple ethnic cultures including the Miao,Dong,and Han,and hosts a number of cultural heritage sites.THE old town of Zhenyua... This ancient city has a history going back over 2,000 years.It has witnessed the fusion of multiple ethnic cultures including the Miao,Dong,and Han,and hosts a number of cultural heritage sites.THE old town of Zhenyuan is located in Zhenyuan County,Qiandongnan Miao and Dong Autonomous Prefecture in southwest China’s Guizhou Province.Covering an area of just around three square kilometers,it sits on the banks of the Wuyang River and is surrounded by mountains.Zhenyuan has been a county for more than 2,281 years and was the prefectural capital during the Yuan(1271-1368)and Qing(1644-1911)dynasties.It is a multicultural region,with Miao,Dong,and other ethnic minorities making up nearly half of the population. 展开更多
关键词 GUIZHOU gateway SOUTHWEST
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The Design of Multi-Connector IoT Central Gateway Based on Raspberry Pi and Docker
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作者 Jie Wang 《Journal of Electronic Research and Application》 2024年第5期66-71,共6页
With the vigorous development of Internet of Things(IoT)technology,the demand for communication and data exchange between different types of IoT devices is increasing day by day.To solve the problems of diversity and ... With the vigorous development of Internet of Things(IoT)technology,the demand for communication and data exchange between different types of IoT devices is increasing day by day.To solve the problems of diversity and complexity of communication protocols between devices,this paper proposes a design scheme of a multi-connector IoT central gateway based on Raspberry Pi and Docker.Through the research and application of related technologies,by integrating multiple communication interfaces and utilizing containerization technology,an efficient,flexible,and scalable IoT central gateway has been realized,which can support the connection and data interaction of multiple communication protocols and provide strong support for the stable operation and development of the IoT system. 展开更多
关键词 Internet of Things Raspberry Pi DOCKER CONNECTOR Central gateway
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Gateway技术构建酵母双杂分析用辣椒cDNA文库 被引量:10
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作者 邱爱连 刘林林 +5 位作者 蔡汉阳 朱万里 黄木坤 陈雄 何水林 张健 《江西农业大学学报》 CAS CSCD 北大核心 2010年第4期791-796,共6页
构建cDNA文库是利用酵母双杂交技术开展特定cDNA分离的重要基础工作。利用Gateway技术,在成功提取辣椒总RNA并分离mRNA的基础上,合成了3种读码框的双链cDNA,通过BP反应构建其相应的入门文库,再通过LR反应将cDNA迅速且定向地重组到酵母... 构建cDNA文库是利用酵母双杂交技术开展特定cDNA分离的重要基础工作。利用Gateway技术,在成功提取辣椒总RNA并分离mRNA的基础上,合成了3种读码框的双链cDNA,通过BP反应构建其相应的入门文库,再通过LR反应将cDNA迅速且定向地重组到酵母双杂分析用目的载体pDESTTM22上,构建成高质量目的文库。经检测入门文库和目的文库滴度均达到(4.0~5.0)×106cfu/mL,文库总容量为(2.4~3.0)×107cfu,平均插入片段大小为1250bp,其质量完全满足于进一步利用酵母双杂体系分析目的基因互作蛋白cDNA。 展开更多
关键词 辣椒 入门文库 目的文库 gateway 酵母双杂
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利用Gateway克隆技术大规模克隆拟南芥转录因子 被引量:19
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作者 梅文倩 宋文强 +2 位作者 潘怡 巩威 朱玉贤 《分子植物育种》 CAS CSCD 2004年第3期358-364,共7页
随着越来越多基因组全序列的测定完成,基因组的研究进入了功能研究阶段。功能基因组的研究需要把大量基因连入不同载体,传统的酶切连接构建载体的方法不能满足这种大规模克隆的需要。Gateway技术是一种高效的大规模克隆系统,而且对载体... 随着越来越多基因组全序列的测定完成,基因组的研究进入了功能研究阶段。功能基因组的研究需要把大量基因连入不同载体,传统的酶切连接构建载体的方法不能满足这种大规模克隆的需要。Gateway技术是一种高效的大规模克隆系统,而且对载体和宿主没有依赖性。本文利用Gateway大规模克隆技术将16个拟南芥转录因子的ORF克隆入植物表达载pPTV和酵母表达载体pYTV中,酵母融合表达实验和West-ern-blot检测证明了该克隆途径的可行性,并且得到的His-Tag融合蛋白,为拟南芥转录因子的大规模蛋白质组学研究奠定了良好的基础,同时基因序列的聚类分析为将来进一步的功能研究提供了有利信息。 展开更多
关键词 gateway克隆技术 拟南芥 转录因子 聚类分析 功能基因组
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基于Gateway技术的低成本植物双分子荧光互补分析系统 被引量:6
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作者 周洁 王栩鸣 +5 位作者 陈斌 陈娟 杨勇 余初浪 严成其 陈剑平 《浙江农业学报》 CSCD 北大核心 2013年第5期1024-1030,共7页
创制基于Gateway重组技术的双分子荧光互补(bimolecular fluorescence complementation,BiFC)载体,同时创制低成本低克隆背景的Gateway入门载体。利用入门载体,采用Gateway技术快速将目标基因重组到BiFC目标载体,并采用农杆菌注射侵染... 创制基于Gateway重组技术的双分子荧光互补(bimolecular fluorescence complementation,BiFC)载体,同时创制低成本低克隆背景的Gateway入门载体。利用入门载体,采用Gateway技术快速将目标基因重组到BiFC目标载体,并采用农杆菌注射侵染烟草的方法观察荧光互作结果。构建的Gateway入门载体,由于带有ccdB基因,非线性化的载体无法在ccdB敏感菌株中生长,因此消除了载体本身的背景。同时由XcmⅠ酶切后产生的T突出,可以很好地与PCR产物结合,连接效率和普通商业化T载体一致。此外,由于载体可由实验室自行制备且采用了TA克隆,创制入门克隆的成本大大降低。借助该研究改造的BiFC目标载体,目的基因能快速地重组到目的载体用于基因互作鉴定。利用本研究构建的Gateway系统,能够以较低成本快速的进行基因克隆和互作研究。 展开更多
关键词 入门载体 BIFC 基因互作 gateway
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使用与Gateway技术兼容的T载体获得入门克隆 被引量:10
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作者 陈其军 安瑞 +2 位作者 周海梦 陈珈 王学臣 《生物化学与生物物理进展》 SCIE CAS CSCD 北大核心 2004年第10期951-954,共4页
与Gateway技术兼容的农杆菌双元载体系统已开始应用于植物功能基因组的研究 ,但应用这些载体系统的一个瓶颈问题 ,是如何简单、经济和高效地将PCR产物或其他来源的目的DNA片段构建到入门载体上获得入门克隆 .为此 ,将传统的TA克隆技术与... 与Gateway技术兼容的农杆菌双元载体系统已开始应用于植物功能基因组的研究 ,但应用这些载体系统的一个瓶颈问题 ,是如何简单、经济和高效地将PCR产物或其他来源的目的DNA片段构建到入门载体上获得入门克隆 .为此 ,将传统的TA克隆技术与Gateway重组克隆技术进行整合 ,构建了与Gateway技术兼容的两种TA克隆载体 ,用于在克隆PCR产物或其他来源的目的DNA片段的同时获得入门克隆 .利用兼容Gateway技术的TA克隆载体有效地解决了上述瓶颈问题 . 展开更多
关键词 gateway克隆技术 TA克隆技术 入门载体 T载体 入门克隆
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Gateway技术构建结缕草低温和干旱诱导cDNA文库及其质量鉴定 被引量:11
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作者 王舟 宗俊勤 +1 位作者 郭海林 刘建秀 《草业科学》 CAS CSCD 北大核心 2012年第6期911-917,共7页
为研究结缕草(Zoysia japonica)胁迫响应信号转导途径中的关键基因,揭示结缕草抗逆分子机制,建立结缕草功能基因组学研究基础平台,以经低温、干旱处理的结缕草为材料,采用Gateway技术构建了首个结缕草低温和干旱诱导的标准cDNA文库。文... 为研究结缕草(Zoysia japonica)胁迫响应信号转导途径中的关键基因,揭示结缕草抗逆分子机制,建立结缕草功能基因组学研究基础平台,以经低温、干旱处理的结缕草为材料,采用Gateway技术构建了首个结缕草低温和干旱诱导的标准cDNA文库。文库质量分析表明,未扩增的原始文库滴度1.76×106 pfu.mL-1,库容7.04×106 pfu,插入片段平均大小大于1kb,重组率90%。文库质量优良,可能包含大量新基因,不仅能为结缕草功能基因组分析提供必要资源,也可为后期进行高通量EST测序、发掘新抗逆相关基因、制作基因芯片等研究奠定基础。 展开更多
关键词 结缕草 CDNA文库 低温胁迫 干旱胁迫 构建 gateway技术
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单子叶植物RNA干扰和过表达Gateway载体的构建 被引量:10
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作者 毕惠惠 王根平 +1 位作者 王成社 夏兰琴 《植物遗传资源学报》 CAS CSCD 北大核心 2013年第1期115-123,共9页
基因枪和农杆菌介导的遗传转化是目前常用的两种单子叶植物遗传转化方法。载体的发展和改良是提高植物遗传转化效率的重要基础,RNA干扰载体和过表达载体是目前通过遗传转化研究植物基因功能的主要工具。Gateway克隆技术是一种基于lambd... 基因枪和农杆菌介导的遗传转化是目前常用的两种单子叶植物遗传转化方法。载体的发展和改良是提高植物遗传转化效率的重要基础,RNA干扰载体和过表达载体是目前通过遗传转化研究植物基因功能的主要工具。Gateway克隆技术是一种基于lambda噬菌体特异位点重组特性的通用克隆技术,该技术可以将大批目的基因方便、快捷地连接到受体载体上。本文利用Gateway技术结合传统酶切、连接方法,构建了适用于单子叶植物基因枪和农杆菌转化的RNA干扰Gateway载体pAHC-PSK-RNAi、pClean-G185-RNAi和过表达Gateway载体pAHC-PSK-OE和pClean-G185-OE,为利用基因枪和农杆菌介导的遗传转化,在小麦和水稻等单子叶植物中进行规模化基因功能研究奠定了基础。 展开更多
关键词 单子叶植物 遗传转化 RNA干扰 过表达 gateway
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Gateway~技术构建交链孢菌JH505 cDNA文库 被引量:12
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作者 龙松华 张宁 +2 位作者 邱德文 黎定军 黄炜 《微生物学报》 CAS CSCD 北大核心 2005年第6期963-965,共3页
Gateway○R技术构建cDNA文库,利用λ噬菌体的位点特异性重组,避免使用限制性内酶切割cDNA,能够解决常规方法构建cDNA文库的技术缺陷。首次应用Gateway○R技术构建交链孢菌cDNA文库,经检测cDNA入门文库的滴度达到1×107cfumL,文库总... Gateway○R技术构建cDNA文库,利用λ噬菌体的位点特异性重组,避免使用限制性内酶切割cDNA,能够解决常规方法构建cDNA文库的技术缺陷。首次应用Gateway○R技术构建交链孢菌cDNA文库,经检测cDNA入门文库的滴度达到1×107cfumL,文库总容量为9×107cfu,平均插入片段为1510bp。通过LR重组把入门文库转换为表达文库,表达文库的滴度为1.58×106cfu/mL,文库总容量为6.32×106cfu,平均插入片段大小为1680bp。表达文库的构建为进一步克隆植物激活蛋白基因打下了基础。 展开更多
关键词 gateway技术 交链孢菌 激活蛋白 cDNA入门文库 CDNA表达文库
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应用Gateway技术构建水稻OsDAD1基因的RNA干涉载体 被引量:16
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作者 徐化学 熊建华 傅彬英 《分子植物育种》 CAS CSCD 2007年第1期133-136,共4页
近年来,RNA干涉广泛应用于植物基因功能研究。Gateway技术是通过采用多重载体高效构建目标基因及任何DNA片段克隆和表达载体的研究平台,该技术以λ噬菌体的位点特异性重组体系为基础,两端具有重组位点的DNA片段或目标基因可以非常容易... 近年来,RNA干涉广泛应用于植物基因功能研究。Gateway技术是通过采用多重载体高效构建目标基因及任何DNA片段克隆和表达载体的研究平台,该技术以λ噬菌体的位点特异性重组体系为基础,两端具有重组位点的DNA片段或目标基因可以非常容易地被重组克隆到含同源重组位点的载体上。这种以重组为基础的克隆体系目前已被广泛应用于分子生物学中来分析基因的功能。本文应用Gateway技术体系替代传统载体构建方法,构建了水稻基因OsDAD1的RNA干涉载体,从而证明了Gateway技术的可行性。 展开更多
关键词 RNA干涉 载体 gateway OsDAD1基因
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利用Gateway技术构建黄瓜HPL基因的RNA干扰载体 被引量:3
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作者 王聪颖 陈书霞 +5 位作者 陈巧 郝丽宁 孟焕文 万旭花 申晓青 程智慧 《西北农业学报》 CAS CSCD 北大核心 2013年第2期152-158,共7页
为探讨黄瓜脂氢过氧化物裂解酶基因(HPL)下调表达对黄瓜果实挥发性醛类物质合成的影响,设计1对特异引物,以黄瓜pDONR201-HPL质粒为模板,通过PCR扩增出323bp的片段。利用基于同源重组原理的Gateway技术,将扩增的片段通过BP反应连接到入... 为探讨黄瓜脂氢过氧化物裂解酶基因(HPL)下调表达对黄瓜果实挥发性醛类物质合成的影响,设计1对特异引物,以黄瓜pDONR201-HPL质粒为模板,通过PCR扩增出323bp的片段。利用基于同源重组原理的Gateway技术,将扩增的片段通过BP反应连接到入门载体pDONR201中。对扩增出的323bp的片段进行测序,测序结果显示该序列与原序列同源性为100%。在此基础上,通过LR反应将目的片段插入过表达的植物RNA干涉载体pHellsgate8,构建黄瓜HPL基因的RNAi表达载体pHellsgate8-HPLi,分别经限制性内切酶XhoⅠ、XbaⅠ酶切后获得323bp的片段,与预期结果一致。采用热击法将pHellsgate8-HPLi转化农杆菌GV3101,经菌液PCR扩增获得323bp的目的条带,表明RNAi表达载体pHellsgate8-HPLi已成功转入农杆菌中。 展开更多
关键词 黄瓜 HPL基因 RNAi干扰 克隆载体 表达载体 gateway技术
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Gateway技术快速构建百合双价抗病毒RNAi表达载体 被引量:5
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作者 徐品三 白建芳 +1 位作者 刘纪文 李焕改 《中国农学通报》 CSCD 北大核心 2011年第4期144-147,共4页
为获得适合转化百合的双价RNAi表达载体,以感病百合叶片的总RNA为模板,分别扩增400bp左右的LSV和LMoVCP基因。通过重叠延伸PCR技术获得长约800bp的LSV-LMoV融合基因。利用Gateway技术,通过BP反应及LR反应将LSV-LMoV融合基因克隆到双元载... 为获得适合转化百合的双价RNAi表达载体,以感病百合叶片的总RNA为模板,分别扩增400bp左右的LSV和LMoVCP基因。通过重叠延伸PCR技术获得长约800bp的LSV-LMoV融合基因。利用Gateway技术,通过BP反应及LR反应将LSV-LMoV融合基因克隆到双元载体pH7GWIWG2(Ⅱ),成功构建了含2种病毒基因的RNAi表达载体pH7GWIWG2(Ⅱ)-LSV-LMoV。将构建好的双价表达载体导入农杆菌EHA105,PCR和测序表明所构建的载体及获得的工程菌与预期的完全一致。 展开更多
关键词 百合无症病毒 百合斑驳病毒 RNAI载体 重叠延伸PCR gateway技术
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