Genetic Design of a Gene for Human Metallothionein II and Its Expression as an Active Fusion Protein in Escherichia coli
スポンサーリンク
概要
- 論文の詳細を見る
A synthetic DNA coding for human metallothionein II (hMT-II) was designed for efficient expression in Escherichia coli and cloned into a vector, pUEX2. Upon induction by heating, the gene for hMT-II was expressed as a fusion protein between β-galactosidase and hMT-II. The fusion protein accounted for approximately 24% of the total protein in the cells and accumulated in the cells as an insoluble form. The fused hMT-II protein with a molecular mass of approximately 122 kDa was purified to homogeneity by washing the inclusion bodies with a solution of Triton X-100. The insoluble protein was easily solubilized in the presence of 2-mercaptoethanol and sodium dodecyl sulfate without any need for denaturants, such as urea or guanidine hydrochloride. The fused hMT-II protein bound to radioactive cadimium. A competition experiment with radioactive cadimium and various heavy methals showed that the hMT-II fusion protein had a higher affinity for Cu^<2+>, Co^<2+>, and Ag^+ ions than Cd^<2+> and Zn^<2+> ions. Thus, we succeeded in over expression of hMT-II as an active form in E. coli.
- 公益社団法人日本生物工学会の論文
- 1994-02-25
著者
-
Murooka Yoshikatsu
Depart. Of Bioteclnology Graduate School Of Eng. Osaka Univ.
-
Kuwata H
Morinaga Milk Ind. Co. Ltd. Kanagawa Jpn
-
Yamashita Mitsuo
Department Of Agricultural Chemistry Faculty Of Bioresources Mie University Department Of Biotechnol
-
KUWATA HIDEFUMI
Department of Fermentation Technology, Faculty of Engineering, Hiroshima University
-
MURAKAMI HIDEHIKO
Department of Fermentation Technology, Faculty of Engineering, Hiroshima University
-
Murakami Hidehiko
Department Of Fermentation Technology Faculty Of Engineering Hiroshima University
関連論文
- Characterization of the C-terminal truncated form of amylopullulanase from Lactobacillus plantarum L137(MICROBIAL PHYSIOLOGY AND BIOTECHNOLOGY)
- Structural Characterization of a New Acidic Ecopolysaccharide and Cyclic (1→2) β-Glucan Produced by Rhizobium huakuii Forming Nodules on Astragalus sinicus
- Characterization of Gene Encoding Amylopullulanase from Plant-Originated Lactic Acid Bacterium, Lactobacillus plantarum L137(MICROBIAL PHYSIOLOGY AND BIOTECHNOLOGY)
- Promotion of Metal Accumulation in Nodule of Astragalus sinicus by the Expression of the Iron-regulated Transporter Gene in Mesorhizobium huakuii subsp. rengei B3(MICROBIAL PHYSIOLOGY AND BIOTECHNOLOGY)
- Production of Vitamin B_ in Genetically Engineered Propionibacterium freudenreichii(MICROBIAL PHYSIOLOGY AND BIOTECHNOLOGY)
- Molecular Analysis of Promoter Elements from Propionibacterium freudenreichii (GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)
- Heterologous Expression of a Gene Encoding Cholesterol Oxidase in Probiotic Strains of Lactobacillus plantarum and Propionibacterium freudenreichii under the Control of Native Promoters
- Development of a Host-Vector System for Lactobacillus plantarum L137 Isolated from a Traditional Fermented Food Produced in the Philippines
- Characterization of the hemB Gene Encoding δ-aminolevulinic Acid Dehydratase from Propionibacterium freudenreichii
- Use of lac Gene Fusions to Study Regulation of Tyramine Oxidase, Which is Involved in Derepression of Latent Arylsulfatase in Escherichia coli
- In Vivo Cloning of Arylsulfatase-Tyramine Oxidase Genes in rec Strains of Klebsiella aerogenes
- Secretory Production of Ascorbate Oxidase by Cultured Cells of Cucumber
- Characterization and Identification of Bacterial Strains Isolated from Corroded Concrete in the Accumulation Stratum and Their Resistance Levels to Heavy Metals
- Isolation and Characterization of a Heavy Metal-Binding Protein from a Heavy Metal-Resistant Strain of Thiobacillus sp.
- Resistance to and Uptake of Heavy Metals in Mushrooms
- 711 Bioremediation system for heavy metals using leguminous plant and Rhizobia symbiosis
- Molecular Cloning and Characterization of Two cDNAs Encoding Asparagine Synthetase from Astragalus sinicus Nodules
- Enterobater colacae A105,Isolated from the Surface of Root Nodules of Astragalus sinicus cv. Japan, Stimulates Nodulation by Rhizobium huakuii bv. renge
- Formation of Root Nudules by Rhizobium huakuii biovar. renge bv. nov. on Astragalus sinicus cv. Japan
- Purification, Characterization, and Crystallization of Monoamine Oxidase from Escherichia coli K-12
- Nucleotide Sequence of the Gene for Monoamine Oxidase (maoA) from Escherichia coli
- Characterization and Sequencing of the Region Required for Replication of a Non-Selftransmissible Plasmid pEC3 Isolated from Erwinia carotovora subsp. carotovora
- Transformation of Dimethyl Sulfide and Related Compounds by Cultures and Cell Extracts of Marine Phytoplankton
- Toward Elucidating the Full Spectrum of Mite Allergens : State of the Art
- Accumulation of Compatible Solutes, Ectoine and Hydroxyectoine, in a Moderate Halophile, Halomonas elengata KS3 Isolated from Dry Salty Land in Thailand
- Variation of the Amino Acid Content of Arabidopsis Seeds by Expressing Soybean Aspartate Aminotransferase Gene
- Isolation, Characterization, and Molecular Cloning of a Thermostable Xylitol Oxidase from Streptomyces sp.IKD472
- Root Hair Deformation of Symbiosis-Deficient Mutants of Lotus japonicus by Application of Nod Factor from Mesorhizobium loti
- Cyclic AMP Regulation of the mob Operon of the Non-Self-Transmissible Plasmid pEC3 Isolated from a Phytopathogenic Bacterium
- Recent Advances in Studies on Streptomyces Cholesterol Oxidase and Gene Clusters Involved in Steroid Catabolism in Prokaryotes
- Genetic Modification of the Streptomyces Cholesterol Oxidase Gene for Expression in Escherichia coli and Development of Promoter-Probe Vectors for Use in Enteric Bacteria
- Purification and Characterization of the 3-Ketosteroid-Δ^1-Dehydrogenase of Arthrobacter simplex Produced in Streptomyces lividans^1
- Secretory Overproduction of Streptomyces Cholesterol Oxidase by Streptomyces lividans with a Multi-Copy Shuttle Vector
- Phylogenetic Position of Mesorhizobium huakuii subsp.rengei, a Symbiont of Astragalus sinicus cv.Japan
- AsNODc22, a Novel Nodulin Gene of Astragalus sinicus, Encodes a Protein that Localizes along the Cell Wall of Bacteria-Infected Cells in a Nodule
- Phylogeny of Bacterial Symbionts of the Laguminous Tree Acacia mangium
- Identification of Cholesterol Oxidase from Fast-Growing Mycobacterial Strains and Rhodococcus sp.
- Origin of Fan Palm (Livistona chinensis R. Br. var. subglobosa Becc.) in Aoshima, Japan
- The Monoamine Regulon Including Syntheses of Arylsulfatase and Monoamine Oxidase in Bacteria
- Heavy Metal Particle Resistance in Thiobacillus intermedius 13-1 Isolated from Corroded Concrete
- Root Hair Deformation of Symbiosis-Deficient Mutants of Lotus japonicus by Application of Nod Factor from Mesorhizobium loti
- Expression of a Major House Dust Mite Allergen Gene from Dermatophagoides farinae in Lotus japonicus Accession Miyakojima MG-20 (Plant Biotechnology)
- Construction of an Additional Metal-Binding Site in Human Metallothionein-2(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
- Interactions of Arsenic with Human Metallothionein-2
- Purification and Characterization of Monoamine Oxidase from Klebsiella aerogenes
- A Simple PCR Method for Distinguishing Saccharomyces cerevisiae from Its Sibling species by Amplification of the RPL2 Region
- Root Hair Abundant Genes LjRH101 and LjRH102 Encode Peroxidase and Xyloglucan Endotransglycosylase in Lotus japonicus(Plant Biotechnology)
- Production of 5-aminolevulinic acid by Propionibacterium acidipropionici TISTR442(MICROBIAL PHYSIOLOGY AND BIOTECHNOLOGY)
- Pollen Development and Tube Growth are Affected in the Symbiotic Mutant of Lotus japonicus, crinkle
- Gene Targeting in a Klebsiella sp. by Fusaric Acid Selection and the Use of Temperature Sensitive Replicon
- Nucleotide Sequences of BstUI Repetitive Elements from Nuclear DNA of Chlorella ellipsoidea C-87
- Rhodobacter sphaeroides Mutants Which Accumulate 5-Aminolevulinic Acid under Aerobic and Dark Conditions
- Purification of the Early Sporulation Gene spo0F Product of Bacillus subtilis(Microbiology & Fermentaion Industry)
- Genetic and Protein Engineering of Diagnostic Enzymes, Cholesterol Oxidase and Xylitol Oxidase
- Adsorption of Bacterial Cells to Crystal Particles of Heavy Metals : Role of Electrostatic Interaction
- Nucleotide Sequence Analysis of a Region Upstream of the Cholesterol Oxidase-Cytochrome P450 Operon of Streptomyces sp. SA-COO Revealing Repeating Units Coding for Putative Transmembrane and DNA-Binding Proteins
- Efficient Transformation of Mesorhizobium huakuii subsp.rengei and Rhizobium Species
- 167 Bioremediation of heavy metals by expression of the metallothionein in Renge-soh nodules
- Characterization of Pseudomonas stutzeri NT-I capable of removing soluble selenium from the aqueous phase under aerobic conditions(ENVIRONMENTAL BIOTECHNOLOGY)
- Lipoteichoic acids on Lactobacillus plantarum cell surfaces correlate with induction of interleukin-12p40 production
- Genetic Design of a Gene for Human Metallothionein II and Its Expression as an Active Fusion Protein in Escherichia coli
- Amino Acid Residues Specific for the Catalytic Action towards α-1,6-Glucosidic Linkages in Klebsiella Pullulanase
- 562 "Guided Genomic" Walking", a Systematic Approach to Clone Intermediate-Sized Restriction fragments Extending Outside the Boundaries of Known Sequences
- Prophylactic effect of Lactobacillus oral vaccine expressing a Japanese cedar pollen allergen(MEDICAL BIOTECHNOLOGY)
- Genetic Manipulation System in Propionibacteria
- Prophylactic effect of Lactobacillus oral vaccine expressing a Japanese cedar pollen allergen
- DNA Synthesis and Fragmentation in Bacteroids during Astragalus sinicus Root Nodule Development