Abstract: The present invention relates to a method for detecting protein-protein interactions in living cells, and more particularly, to a method for providing cells comprising a first construct and a second construct, wherein the first construct comprises a polynucleotide encoding a first fusion protein which comprises a bait protein, a first fluorescent protein and a CBD (cellulose-binding domain) protein, and wherein the second construct comprises a polynucleotide encoding a second fusion protein which comprises a prey protein and a second fluorescent protein so as to allow formation of inclusion bodies, and detecting interactions between the bait protein and the prey protein that are displayed by inclusion bodies, a method for isolating the prey protein bound to the bait protein using the cells comprising the constructs, the cells, and a kit for detecting protein-protein interactions, comprising the constructs.

Abstract: The present invention relates to a metagenome-derived alkaline phosphatase, and more particularly to a novel, metagenome-derived alkaline phosphatase screened using a artificial genetic circuit that detects phenolic compounds, and a preparation method thereof. A novel alkaline phosphatase according to the present invention has high activity of dephosphorylating DNA and can be inactivated rapidly by simple heat treatment. Thus, it can be used for a dephosphorylation reaction so that genetic manipulations, including genetic cloning, become efficient. In addition, it can be actively expressed in recombinant microorganisms, and thus can be used in various assays, including enzyme immunoassay.

Abstract: A method of detecting and quantifying various enzymatic activities using a constructed artificial genetic circuit GESS (genetic enzyme screening system) for sensing phenolic compounds and a method of screening a trace of activities of target enzymes from a metagenome using the artificial genetic circuit, thereby securing target enzyme genes. When the method for screening and quantifying target enzymatic activity is used, useful genes can be screened from various genetic communities, including environmental or metagenomic libraries, at a single cell level in high throughput (million/day). Further, the sensitivity of the genetic circuit to phenol derivatives and the expression thereof can be controlled, and thus the genetic circuit can rapidly sense and quantify various enzymatic activities. Thus, the method can be advantageously used in the protein engineering technology for enzyme modification.

Abstract: The present application relates to a method for detecting ligand using a biosensor applied the FRET(fluorescence resonance energy transfer) phenomenon. More particularly, the method may be used for simply detecting a ligand in a sample by measuring the FRET of a biosensor under the conditions in which a specific critical temperature is maintained. The method may use a phenomenon in which a ligand-binding protein in a biosensor shows reversible unfolding at a temperature higher than the specific critical temperature and the level of the unfolding changes depending on the concentration of a ligand. The method can be widely applied to a variety of kinds of FRET biosensors using the ligand-binding protein.

Abstract: A method for cloning and expressing a target gene by homologous recombination, and more particularly a method for cloning and expressing a target gene by homologous recombination, wherein a host cell transformed with a recombinant vector and a plasmid containing a recombinase system is introduced with a linear DNA fragment comprising a target gene and a sequence having homology to the recombinant vector. Because complicated genetic engineering steps, such as the restriction enzyme treatment and ligation of a vector and a target gene, are not required, the cloning of a gene can be performed without needing a high degree of skill, and enzyme cost can be reduced. The inventive method can be effectively used for the massive, high-speed cloning and protein expression of genes, and the disclosed pRMT-iTGR system can be used as an analytical means for improving high-efficiency recombinase.

Abstract: The present invention relates to methods for dynamically detecting the interactions between various materials including bioactive molecules and for detecting target molecules. More specifically, the present invention relates to a method for dynamically detecting bait-prey interactions and a method for easily detecting target molecules which blocks or activates the interactions, the method comprising: allowing a material capable of forming a nano-assembly matrix, a bait and a prey to interact with each other, and analyzing whether a nano-assembly matrix is formed by the interaction between the bait and the prey in vitro or in vivo; or allowing a material capable of forming a nano-assembly matrix, a bait and a prey to interact with each other, inducing the formation of a nano-assembly matrix by a mediator (regulator) material in vitro or in vivo, and then analyzing whether the prey and the bait co-localizes on the nano-assembly matrix.

Abstract: A method for cloning and expressing a target gene by homologous recombination, and more particularly a method for cloning and expressing a target gene by homologous recombination, wherein a host cell transformed with a recombinant vector and a plasmid containing a recombinase system is introduced with a linear DNA fragment comprising a target gene and a sequence having homology to the recombinant vector. Because complicated genetic engineering steps, such as the restriction enzyme treatment and ligation of a vector and a target gene, are not required, the cloning of a gene can be performed without needing a high degree of skill, and enzyme cost can be reduced. The inventive method can be effectively used for the massive, high-speed cloning and protein expression of genes, and the disclosed pRMT-iTGR system can be used as an analytical means for improving high-efficiency recombinase.

Abstract: To provide a promoter, a recombinant DNA, a gene expression vector, an expression vector, and a transformant, which are capable of expressing a gene without inducing gene expression with an inducer; and a method for producing a protein and a kit therefor, which can be operated easily and performed inexpensively by convenient and inexpensive steps.

Abstract: A protein biosensor having increased signal intensity to sugar concentration and the use thereof. A fluorescent indicator protein is described, in which fluorescent proteins of different emission ranges are fused to both ends of a binding protein undergoing a conformational change due to binding of sugars, such that a change in the concentration of various sugars involved in intracellular metabolism, e.g., maltose, can be detected by measuring a change in emission intensities, caused by FRET (fluorescence resonance energy transfer). A method for detecting a change in the concentration of sugars using such fluorescent indicator protein is also described. The fluorescent indicator protein has excellent signal intensity enabling precision measurement of intracellular concentration of various sugars. In one implementation, high signal intensity fluorescent indicator proteins can be prepared from proteins otherwise used as biosensors, to enable them to be more widely useful.

Abstract: The present invention relates to a novel obligately symbiotic thermophile Symbiobacterium toebii SC-1 (Accession NO: KCTC 0685BP), and a thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase produced by Symbiobacterium toebii SC-1. The present invention also relates to a pure culture method of Symbiobacterium toebii SC-1, a method of growth measurement for Symbiobacterium toebii SC-1 and its relative symbiotic bacteria showing low-growth yield using nitrate respiration, and a screening method of relative symbiotic strains of Symbiobacterium toebii SC-1 from the environment using a specific antibody to it. Therefore, the thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase produced by Symbiobacterium toebii SC-1 in this invention can provide more stable catalysts in enzymatic biotransformation processes which enzymatically produce valuable medico-amino acids 3,4-dehydroxy-L-phenylalanine (L-DOPA) and L-tryptophan.

Abstract: A method for cloning and expressing a target gene by homologous recombination, and more particularly a method for cloning and expressing a target gene by homologous recombination, wherein a host cell transformed with a recombinant vector and a plasmid containing a recombinase system is introduced with a linear DNA fragment comprising a target gene and a sequence having homology to the recombinant vector. Because complicated genetic engineering steps, such as the restriction enzyme treatment and ligation of a vector and a target gene, are not required, the cloning of a gene can be performed without needing a high degree of skill, and enzyme cost can be reduced. The inventive method can be effectively used for the massive, high-speed cloning and protein expression of genes, and the disclosed pRMT-iTGR system can be used as an analytical means for improving high-efficiency recombinase.

Abstract: To provide a promoter, a recombinant DNA, a gene expression vector, an expression vector, and a transformant, which are capable of expressing a gene without inducing gene expression with an inducer; and a method for producing a protein and a kit therefor, which can be operated easily and performed inexpensively by convenient and inexpensive steps.

Abstract: To provide a promoter, a recombinant DNA, a gene expression vector, an expression vector, and a transformant, which are capable of expressing a gene without inducing gene expression with an inducer; and a method for producing a protein and a kit therefor, which can be operated easily and performed inexpensively by convenient and inexpensive steps.

Abstract: A protein biosensor having increased signal intensity to sugar concentration and the use thereof. A fluorescent indicator protein is described, in which fluorescent proteins of different emission ranges are fused to both ends of a binding protein undergoing a confornational change due to binding of sugars, such that a change in the concentration of various sugars involved in intracellular metabolism, e.g., maltose, can be detected by measuring a change in emission intensities, caused by FRET (fluorescence resonance energy transfer). A method for detecting a change in the concentration of sugars using such fluorescent indicator protein is also described. The fluorescent indicator protein has excellent signal intensity enabling precision measurement of intracellular concentration of various sugars. In one implementation, high signal intensity fluorescent indicator proteins can be prepared from proteins otherwise used as biosensors, to enable them to be more widely useful.

Abstract: An antibiotic-independent vector for stable vector maintenance and high protein expression and a method for gene expression using this vector are disclosed. The stable maintenance of the vector is due to expression of a glutamic acid racemase to complement a D-glutamic acid auxotroph. No antibiotics, such as ampicillin, are necessary for the stable maintenance of this vector.

Abstract: The present invention relates to a novel obligately symbiotic thermophile Symbiobacterium toebii SC-1(Accession NO: KCTC 0685BP), and a thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase produced by Symbiobacterium toebii SC-1. The present invention also relates to a pure culture method of Symbiobacterium toebii SC-1, a method of growth measurement for Symbiobacterium toebu SC-1 and its relative symbiotic bacteria showing low-growth yield using nitrate respiration, and a screening method of relative symbiotic strains of Symbiobacterium toebii SC-1 from the environment using a specific antibody to it. Therefore, the thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase produced by Symbiobacterium toebii SC-1 in this invention can provide more stable catalysts in enzymatic biotransformation processes which enzymatically produce valuable medico-amino acids 3,4-dehydroxy-L-phenylalanine (L-DOPA) and L-tryptophan.

Abstract: The present invention relates to a novel obligately symbiotic thermophile Symbiobacterium toebii SC-1 (Accession NO: KCTC 0685BP), and a thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase produced by Symbiobacterium toebii SC-1. The present invention also relates to a pure culture method of Symbiobacterium toebii SC-1, a method of growth measurement for Symbiobacterium toebii SC-1 and its relative symbiotic bacteria showing low-growth yield using nitrate respiration, and a screening method of relative symbiotic strains of Symbiobacterium toebii SC-1 from the environment using a specific antibody to it. Therefore, the thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase produced by Symbiobacterium toebii SC-1 in this invention can provide more stable catalysts in enzymatic biotransformation processes which enzymatically produce valuable medico-amino acids 3,4-dehydroxy-L-phenylalanine (L-DOPA) and L-tryptophan.

Abstract: To provide a promoter, a recombinant DNA, a gene expression vector, an expression vector, and a transformant, which are capable of expressing a gene without inducing gene expression with an inducer; and a method for producing a protein and a kit therefor, which can be operated easily and performed inexpensively by convenient and inexpensive steps.

Abstract: The present invention relates to an antibiotic-independent vector for constant high-expression and a method for gene expression using this vector. The stable expression of a recombinant gene is maintained by a stable plasmid through the use of genes essential for the growth and proliferation of the microorganism as a marker, instead of antibiotics, such as ampicillin.