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The Development of protein microarrays and Their Applications in DNA-protein and protein-protein Interaction Analyses of Arabidopsis Transcription Factors

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Molecular Plant 2008年第1期1卷 27-41页

作者： Wei Gong Kun He Mike Covington S. R Dinesh-Kumar Michael Snyder Stacey L. Harmer Yu-Xian Zhu Xing Wang Denga Peking-Yale Joint Center for Plant Molecular Genetics and Agrobiotechnology Co and Plant Genetic Engineering Peking University Beijing 100871 China Department of Molecular Cellular and Developmental Biology Yale University New Haven Connecticut 06520 USA Section of Plant Biology College of Biological Sciences University of California Davis 95616 USA

We used our collection of Arabidopsis transcription factor （TF） ORFeome clones to construct protein microarrays containing as many as 802 TF proteins. These protein microarrays were used for both protein-DNA and proteinprotein interaction analyses. For protein-DNA interaction studies, we examined AP2/ERF family TFs and their cognate cis-elements. By careful comparison of the DNA-binding specificity of 13 TFs on the protein microarray with previous non-microarray data, we showed that protein microarrays provide an efficient and high throughput tool for genome-wide analysis of TF-DNA interactions. This microarray protein-DNA interaction analysis allowed us to derive a comprehensive view of DNA-binding profiles of AP2/ERF family proteins in Arabidopsis. It also revealed four TFs that bound the EE （evening element） and had the expected phased gene expression under clock-regulation, thus providing a basis for further functional analysis of their roles in clock regulation of gene expression. We also developed procedures for detecting protein interactions using this TF protein microarray and discovered four novel partners that interact with HY5, which can be validated by yeast two-hybrid assays. Thus, plant TF protein microarrays offer an attractive high-throughput alternative to traditional techniques for TF functional characterization on a global scale.

关键词： protein microarrays PLANT physiology proteinS PLANT genetics ARABIDOPSIS thaliana TRANSCRIPTION factors

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Label-free detection repeatability of protein microarrays by oblique-incidence reflectivity difference method

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Science China(Physics,Mechanics & Astronomy) 2012年第12期55卷 2347-2350页

作者： DAI Jun LI Lin WANG JingYi HE LiPing LU HuaBin RUAN KangCheng JIN KuiJuan YANG GuoZhen Beijing National Laboratory for Condensed Matter Physics Institute of Physics Chinese Academy of Sciences Beijing 100190 China State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

We examine the repeatabilities of oblique-incidence reflectivity difference(OIRD) method for label-free detecting biological molecular interaction using protein *** experimental results show that the repeatabilities are the same in a given microarray or microarray-microarray and are consistent,indicating that OIRD is a promising label-free detection technique for biological microarrays.

关键词： oblique-incidence reflectivity difference(OIRD) protein microarrays specificity repeatability

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Detection of hybridization of protein microarrays using an oblique-incidence reflectivity difference method

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Science China(Physics,Mechanics & Astronomy) 2010年第7期53卷 1230-1233页

作者： LU Heng WEN Juan WANG Xu YUAN Kun LU HuiBin ZHOU YueLiang JIN Kui-Juan YANG GuoZhen LI Wei RUAN KangCheng Beijing National Laboratory for Condensed Matter Physics Institute of PhysicsChinese Academy of SciencesBeijing 100190China Key Laboratory of Proteomics Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China

Mouse-Immunoglobulin G(mouse-IgG) with different concentrations in a range from 1000 to 0.0128 μg/mL and a specific hybridization with goat anti-mouse IgG were detected successfully by using an oblique-incidence reflectivity difference(OI-RD) *** detection signals,consisting of an imaginary part(Im{Δp-Δs}) and a real part(Re{Δp-Δs}) of OI-RD,were obtained *** detection results of hybridization by OI-RD were in accord with that of traditional fluorescent *** particular,we label-freely detected the washed mouse-IgG microarray with a series of concentrations and acquired a linear correlation between OI-RD intensities and the protein concentrations in logarithmic *** detection sensitivity of OI-RD can reach 14 *** experimental results suggest that the OI-RD method has potential applications in proteomics and clinical diagnosis.

关键词： protein microarrays oblique-incidence reflectivity difference(OI-RD) label-free detection

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Label-Free and High-Throughput Detection of protein microarrays by Oblique-Incidence Reflectivity Difference Method

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Chinese Physics Letters 2010年第10期27卷 198-201页

作者：王旭陆珩温娟原昆吕惠宾金奎娟周岳亮杨国桢 Beijing National Laboratory for Condensed Matter Physics Institute of Physics Chinese Academy of Sciences Beijing 100190 School of Science Jiangnan University Wuxi 214122

We label-free detected the biological process of preparing a microarray that includes 400 spots of mouse immunoglobulin G （IgG） as well as the specific hybridization between mouse IgG and goat anti-mouse IgG by an oblique-incidence reflectivity difference （OI-RD） method. The detection results after each process including printing, washing, blocking, and hybridization, demonstrate that the OI-RD method can trace the preparation process of a microarray and detect the specific hybridization between antigens and antibodies. OI-RD is a promising method for label-free and high-throughput detection of biological microarrays.

关键词： protein microarrays REFLECTANCE IMMUNOGLOBULIN G FINITE differences BIOSENSORS MICE as laboratory animals

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A Putative Nuclear CLE-Like （CLEL） Peptide Precursor Regulates Root Growth in Arabidopsis

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Molecular Plant 2012年第4期5卷 955-957页

作者： Ling Meng Bob B. Buchanan Lewis J. Feldman Sheng Luan Department of Plant and Microbial Biology University of California Berkeley CA USA

Dear Editor, It is well established that local intercellular signaling net- works control cell identity and activity. Secreted peptides have been found to play a central role in this capacity in plants （Matsubayashi and Sakagami, 2006; Meng et al., 2010; Meng, 2012）. CLEL, which is the term for CLE （CLV3/ESR-related）-Iike peptides （also previously named as RGFs for root growth fac- tors by Matsuzaki et al. （2010））, regulate root growth pattern and lateral root development in Arabidopsis （Matsuzaki et al., 2010; Meng et al., 2012）. CLEL peptide precursors contain an N-terminal signal peptide （hydrophobic patch）, a diverse vari- able region,

关键词： GENE expression in plants GREEN fluorescent protein SEQUENCE alignment (Bioinformatics) AMINO acid sequence ROOTS (Botany) -- Growth ARABIDOPSIS thaliana protein microarrays PHOTOGRAPHS

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protein microarray platform based on antibody fragments and its applications in simultaneous multianalysis

Protein microarray platform based on antibody fragments and ...

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中国化学会第十三届有机分析与生物分析学术会议

作者： SONG Shi-ping , LI Bin, WU Hai-ping, HU Jun, LI Min-qian, FAN Chun-hai ( Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800 Bio- X Life Science Research Center, Shanghai Jiaotong University", Shanghai 201800)

＜正＞Antibody microarrays have shown great potential for measurement of either a spectrum of target proteins in proteomics or disease-associated antigens in molecular diagnostics. Despite its importance, the applications of antibody microarrays are still limited by a variety of fundamental problems. Among them, cross-linking interferences significantly limits the multiplexing ability in parallel sandwich immunoassays. As a result, it is highly

关键词： protein microarrays Antibody fragments Tumor markers Simultaneous multianalysis

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