Stabilization of Affinity-Tagged Recombinant Protein during/after Its Production in a Cell-Free System Using Wheat-Germ Extract

スポンサーリンク

概要

• 論文の詳細を見る

• We found that the affinity tag fused to the carboxyl (C-) terminal of a single-chain Fv (scFv) antibody was proteolytically degraded in a wheat germ cell-free protein synthesis system. The addition of two extra residues of glycine to the tail of the cMyc tag significantly increased the stability of the tag, suggesting that wheat endogenous carboxypeptidase(s) play a primary role in the C-terminal tag-specific degradation. In addition to the modification of the tag sequence, addition of diisopropyl fluorophosphate, which is known as an inhibitor of carboxypeptidases, prevented the cMyc tag sequence degradation. The effects of other protease inhibitors on the translation reaction and stability of the synthesized protein are also reported.

• 2003-03-25

著者

• Nakano Hideo

  Laboratory of Molecular Biotechnology, Graduate School of Bioagricultural Sciences, Nagoya Universit

• KAWARASAKI YASUAKI

  Laboratory of Molecular Biotechnology, Division of Molecular and Cellular Mechanisms, graduate Schoo

• YAMADA YASUHIRO

  Laboratory of Molecular Biotechnology, Division of Molecular and Cellular Mechanisms, graduate Schoo

• ICHIMORI MAKI

  Laboratory of Molecular Biotechnology, Division of Molecular and Cellular Mechanisms, graduate Schoo

• SHINBATA TOMOYA

  Nippon Flour Mills, Co. Ltd.

• KOHDA KATSUNORI

  Toyota Central R&D Laboratories Inc.

• YAMANE TSUNEO

  Laboratory of Molecular Biotechnology, Division of Molecular and Cellular Mechanisms, graduate Schoo

• Yamane T

  Laboratory Of Molecular Biotechnology Graduate School Of Bioagricultural Sciences Nagoya University:

• Yamane Tsuneo

  Laboraotory Of Bioreaction Engineering Department Of Food Science And Technology Faculty Of Agricult

• Yamane Tsuneo

  Laboratory Of Bioreaction Engineering Department Of Food Science And Technology School Of Agricultur

• Ichimori Maki

  Laboratory Of Molecular Biotechnology Division Of Molecular And Cellular Mechanisms Graduate School

• Shinbata Tomoya

  Nippon Flour Mills Co. Ltd.

• Kohda K

  Toyota Central R&d Laboratories Inc.

• Yamada Y

  Department Of Biotechnology Graduate School Of Engineering Osaka University

• Yamane T

  Laboratory Of Molecular Biotechnology Graduate School Of Biological & Agricultural Sciences Nago

• Yamane T

  Laboratory Of Bioreaction Engineering Department Of Food Science And Technology School Of Agricultur

• Kobayashi K

  Laboratory Of Molecular Biotechnology Graduate School Of Biological & Agricultural Sciences Nago

• Kawarasaki Yasuaki

  Laboratory Of Molecular Biotechnology Division Of Molecular And Cellular Mechanisms Graduate School

• Nakano Hideo

  Laboratory Of Molecular Biotechnology Graduate School Of Bioagricultural Sciences Nagoya University

• Yabuki Takashi

  Protein Research Group Riken Genomic Sciences Center

• Yamane Tsuneo

  Dept. Of Food Science And Technology Faculty Of Agriculture Nagoya University

• Yamane Tsuneo

  Laboratory Of Molecular Biotechnology Graduate School Of Bio- And Agro-sciences Nagoya University

• Yamada Yasuhiro

  Laboratory Of Molecular Biotechnology Division Of Molecular And Cellular Mechanisms Graduate School

• Nakano Hideo

  Laboratory Of Molecular Biotechnology Department Of Bioengineering Sciences Graduate School Of Bioag

• Nakano Hideo

  Department Of Bioengineering Sciences Graduate School Of Bioagricultural Sciences Nagoya University

• KOHDA KATSUNORI

  Toyota Central R&D Laboratories Inc.

関連論文

• High-throughput screening of DNA binding sites for transcription factor AmyR from Aspergillus nidulans using DNA beads display system(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)
• Directed evolution of angiotensin II-inhibiting peptides using a microbead display(METHODS)
• In vitro generation of anti-hepatitis B monoclonal antibodies from a single plasma cell using single-cell RT-PCR and cell-free protein synthesis(MEDICAL BIOTECHNOLOGY)
• Stabilization of Affinity-Tagged Recombinant Protein during/after Its Production in a Cell-Free System Using Wheat-Germ Extract
• 生体硬組織のエキシマレーザー蒸散機構
• レーザー核融合炉用大型プラスチックシェルの製作
• Directed evolution of angiotensin II-inhibiting peptides using a microbead display
• Synthesis of Phospholipids Containing Polyunsaturated Fatty Acids by Phospholipase A2-mediated Esterification with Food-compatible Reagents
• 紫外ファイバーの加熱によるレーザー誘起欠陥除去
• 紫外中空導光路の伝送率向上
• レーザー内視鏡の機能開発
• エキシマレーザーメス用中空導光路
• 紫外ファイバー伝送率の温度緩和特性
• 紫外レーザー伝送中空導光路の改善
• 紫外ファイバーのArFエキシマレーザー伝送限界
• ArFエキシマレーザーと紫外ファイバーのクラッド材質・屈折率構造
• LD励起1064nm Nd : YAGジグザグスラブレーザー増幅器のビーム品質測定
• LD励起8パス構成1064-nmNd : YAGジグザグスラブレーザー増幅器の高平均出力化
• In Vitro Construction and Screening of a Burkholderia cepacia Lipase Library Using Single-Molecule PCR and Cell-Free Protein Synthesis(METHODS)
• Single-Step Single-Molecule PCR or DNA with a Homo-Priming Sequence Using a Single Primer and Hot-Startable DNA Polymerase
• 1B17-3 A Novel Compact Disk-like Microfluidic Device with Micropillars for Enzyme-linked Immunosorbent Assay
• 755 Monitoring of lipase-catalyzed transesterification between eicosapentaenoic acid ethyl ester and tricaprylin by silver ion high performance liquid chromatography and high temperature gas chromatography
• Syntheses of 2-[(1S,3S)-1-Amino-3-carboxy-3-hydroxypropyl]-thiazole-4-carboxylic Acid and the Tripeptide Skeleton of Nosiheptide Containing the Acid
• P-405 SITE-SPECIFIC FUNCTIONALIZATION OF PROTEINS BY ORGANOPALLADIUM REACTIONS
• Improvements in the Cell-Free Production of Functional Antibodies Using Cell Extract from Protease-Deficient Escherichia coli Mutant (Methods)
• Generation of Monoclonal Antibodies Using Simplified Single-Cell Reverse Transcription-Polymerase Chain Reaction and Cell-Free Protein Synthesis(METHODS)
• Active Lactonizing Lipase (LipL) Efficiently Overproduced by Pseudomonas Strains as Heterologous Expression Hosts(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
• Modulator-Mediated Synthesis of Active Lipase of Pseudomonas sp.109 by Escherichia coli Cell-Free Coupled Transcription/Translation System
• Purification and Characterization of a Novel Extracellular Lipase Catalyzing Hydrolysis of Oleyl Benzoate from Acinetobacter nov. sp. Strain KM109
• Increased Production of Lactonizing Lipase (LipL) from Pseudomonas sp. strain 109 by Lipids and Detergents
• Requirement in trans of the Downstream limL Gene for Activation of Lactonizing Lipase form Pseudomonas sp. 109
• 高アスペクト比レーザー核融合用プラスチックシェルの開発
• Accumulation of Translational Inhibitor durig Multi-Hour Cell-Free Protein Synthesis Reaction Using Rabbit Reticulocyte Lysate
• High Speed Polymerase Chain Reaction in Constant Flow
• Roles of Aggrecan, a Large Chondroitin Sulfate Proteoglycan, in Cartilage Structure and Function
• 448 In vitro analysis of the roles of a disulfide bridge and a calcium binding site in Pseudomonas sp. KWI-56 lipase using PCR and Escherichia coli coupled transcription/translation
• レーザーアブレーションを用いた透明電極用TiNの反射防止膜形成
• 生体脂質における紫外レーザー蒸散能の実験的考察
• Increasing Productivity by Removing Choline in Conversion of Phosphatidylcholine to Phosphatidylserine by Phospholipase D
• 573 Increased Productivity by Removing Choline in Conversion of Phosphatidyl-choline to Phosphatidylserine by Phospolipase D.
• 515 Increasing Phosphatidylcholine Content of Lecithin by Phospholipase D
• Phenomenological Background and Some Preliminary Trials of Automated Substrate Supply in pH-Stat Modal Fed-Batch Culture Using a Setpoint of High Limit
• L-Serine Production from Methanol and Glycine with an Immobilized Methylotroph
• Economic Significance of Increasing Cell Mass Concentration for Metabolite Production : A Theoretical Study
• Effect and Control of Glucose Feeding on Bacitracin Production by Fed-Batch Culture of Bacillus licheniformis
• Production of L-Serine from Methanol and Glycine by Resting Cells of a Methylotroph under Automatically Controlled Conditions
• Control of Carbon Source Supply and Dissolved Oxygen by Use of Carbon Dioxide Concentration of Exhaust Gas in Fed-Batch Culture
• Detection and Automatic Control of Ammonium Ion Concentration in Microbial Culture with an Ammonium Ion Selective Electrode
• 生体硬組織の紫外レーザーアブレーション機構
• 565 Charcteristics of Batch Culture of a Recombinant ___- ___- Producing a Foreign Esterase
• Comprison of Cell Productivities among Fed-Batch, Repeated Fed-Batch and Continuous Cultures at High Cell Concentration
• Inhibition and Inactivation of Lipase by Fat Peroxide in the Course of Batch and Continuous Glycerolyses of Fat by Lipase(Biological Chemistry)
• Selective Hydrolysis of Fish Oil by Lipase to Concentrate n-3 Polyunsaturated Fatty Acids(Biological Chemistry)
• Purification and Some Characteristics of Extracellular Lipase from Fusarium oxysporum f.sp.lini
• In Vitro Selection of DNA Binding Sites for Transcription Factor, PhaR, from Paracoccus denitrificans Using Genetic Library on Microbeads and Flow Cytometry(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)
• Glutathione as an Essential Factor for Chaperon-Mediated Activation of Lactonizing Lipase(LipL)from Pseudomonas sp.109
• Er : YAGレーザーと生体硬組織
• レーザー照射における生体硬組織の組成変化
• 紫外レーザーと蛋白質の相互作用
• パルスレーザーを用いたTiN透明電極膜の作成
• レーザーアブレーション法による可視発光アルミナ薄膜の作製
• エキシマレーザー照射によるプラスチックシェルの燃料貯蔵性改善
• 生体脂肪におけるエキシマレーザーアブレーションの高速イメージング
• エキシマレーザー照射による生体硬組織の微細構造変化
• アミノ酸に対するエキシマレーザー照射効果
• 生体硬組織のエキシマレーザーによるアブレーション機構の解明
• アルミナ基板上へのハイドロキシアパタイト形成とその結晶性
• エキシマレーザーによるTiN薄膜の形成
• 燃料ガス保持型プラスチックシェルの開発
• フッ化ビニリデン-3フッ化エチレン共重合体の電気伝導と分子構造
• 1F14-1 Enzymatic preparation of enantiomerically pure 2,3-diacylglycerol
• 2J10-5 Direct separation of regio- and enantiomeric isomers of diacylglycerols by a tandem column high-performance liquid chromatography
• Expression Profiles of Polyhydroxyalkanoate Synthesis-Related Genes in Paracoccus denitrificans
• Cloning and in Vitro and in Vivo Expression of Plant Glutathione S-Transferase Zeta Class Genes(GENETICS,MOLECULAR BIOLOGY,AND GENE ENGINEERING)
• Reduction of Protein Degradation by Use of Protease-Deficient Mutants in Cell-Free Protein Synthesis System of Escherichia coli
• Dosage Effect of Minor Arginyl- and Isoleucy1-tRNAs on Protein Synthesis in an Escherichia coli In Vitro Coupled Transcription/Translation System
• Importance of Disulfide Bridge Formation on Folding of Phospholipase D from Streptomyces antibioticus
• Continuous Production of Phospholipase D by Streptomyces lydicus D-121 Immobilized with Cross-Linked Chitosan Beads
• Acquisition of Data from On-Line Laser Turbidimeter and Calulation of Some Kinetic Variables in Computer-Coupled Automated Fed-Batch Culture
• Extracellular Production of Phospholipase D of Streptomyces antibioticus Using Recombinat Escherichia coli
• Prolonged Cell-free Protein Synthesis in a Batch System Using Wheat Germ Extract
• Phosphatidylglycerol Synthesis by Phospholipase D in a Microporous Membrane Bioreactor : Studies on Enzymatic Conversion of Phospholipids (I)
• 107 Effective Extracellular Production of Bacillus stearothermophilus Esterase by pH-stat modal Fed-batch Culture of Recombinant Bacillus brevis
• An Increased Rate of Cell-free Protein Synthesis by Condensing Wheat-germ Extract with Ultrafiltration Membranes
• Immobilization of Yeast Cells with Cross-Linked Chitosan Beads
• Site-specific incorporation of 4-lodo-L-phenylalanine through opal suppression
• Three Automated Feeding Strategies of Natural Complex Nutrients Utilizing On-Line Turbidity Values in Fed-Batch Culture : A Case Study on the Cultivation of a Marine Microorganism
• Interesterification of Fats and Oils by Immobilized Fungus at Constant Water Concentration
• Kinetic Studies on the Interesterification of Oils and Fats Using Dried Cells of Fungus
• Screening for Fungi That Have High Lipolytic and Acidolytic Activities in Biomass Support Particles
• Avoidance of Solidification of Sesame Oil at Low Temperature by Self-interesterification with Immobilized Lipase
• Purification and Characterization of Nicotinic Acid Dehydrogenase from Pseudomonas fluorescens TN5
• Microbial Production of 6-Hydroxynicotinic Acid, an Important Building Block for the Synthesis of Modern Insecticides
• Production of 6-Hydroxynicotinic Acid from Nicotinic Acid by Resting Cells of Pseudomonas fluorescens TN5
• A Bacillus stearothermophilus Esterase Produced by a Recombinant Bacillus brevis Stabilized by Sulfhydryl Compounds
• 316. Purification and Characterization of a Thermostable Bacillus stearothermophilus Esterase Excreted from a Recombinant Bacillus brevis
• Purification and Properties of a Thermostable Esterase of Bacillus stearothermophilus Produced by Recombinant Bacillus brevis
• Replica Technique for Screening Poly(3-Hydroxyalkanoic Acid)-Producing Bacteria by Nile Blue Staining
• Some Characteristics of Continuous Glyceride Synthesis by Lipase in a Microporous Hydrophobic Membrane Bioreactor
• Continuous Hydrolysis of Olive Oil by Lipase in Microporous Hydrophobic Hollow Fiber Bioreactor

もっと見る 閉じる

スポンサーリンク