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34 results about "Chaperone Gene" patented technology
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Genes that encode cytoplasmic proteins of both prokaryotes and eukaryotes that bind to nascent or unfolded polypeptides and ensure correct folding or transport. Chaperone proteins do not covalently bind to their targets and do not form part of the finished product. Heat-shock proteins are an important sub set of chaperones. Three major families are recognised, the chaperonins (groEL and hsp60), the hsp70 family and the hsp90 family. Outside these major families are other proteins with similar functions including nucleoplasmin, secB and T-cell receptor associated protein.
Remedy for diseases associated with immunoglobulin gene translocation
InactiveUS20070004674A1Easy to produceEasy to implementBiocideOrganic chemistryHSP90 Heat-Shock ProteinsBULK ACTIVE INGREDIENT
A medicament comprising a compound having an inhibitory action on Hsp90 or a pharmaceutically acceptable salt thereof as an active ingredient is used as a therapeutic agent for diseases associated with immunoglobulin gene translocations. In diseases associated with immunoglobulin gene translocations, abnormal enhancement of the expression of a partner gene in immunoglobulin gene translocation participates in the development of the diseases and the progress of the symptoms. By administering the compound having an inhibitory action on Hsp90 or a pharmaceutically acceptable salt thereof, degradation of protein encoded by the partner gene is promoted and the diseases can be treated.
Owner:KYOWA HAKKO KIRIN CO LTD
Method for preparing recombinant human nerve growth factor by using Escherichia coli expression system
ActiveCN102839182AHigh bioactivityEasy to prepareFermentationVector-based foreign material introductionEscherichia coliInclusion bodies
The invention discloses a method preparing a recombinant human nerve growth factor by using an Escherichia coli expression system. Beta-subunit genes of the recombinant human nerve growth factor comprise 6 histidine purification sites, an enterokinase cleavage site and a beta-subunit gene encoding mature peptide of modified human nerve growth factor and a molecular chaperone gene. The beta-subunit genes of the recombinant human nerve growth factor have high expression quantity in the Escherichia coli; generated inclusion body protein has high-efficiency renaturation under the assist of the molecular chaperone; and the recombinant human nerve growth factor obtained by nickel column affinity chromatography after the molecular chaperone is cleavaged accurately by the enterokinase, has a purity greater than 99% and biological activity greater than 500,000 AU / mg.
Owner:WUHAN HITECK BIOLOGICAL PHARMA
Gene fusion mutation library construction method, gene fusion mutation detection method, gene fusion mutation detection device, equipment and storage medium
PendingCN111321202AImprove economyAccurate expression of magnitudeMicrobiological testing/measurementProteomicsA-DNACore gene
The present invention relates to a gene fusion mutation library construction method, a gene fusion mutation detection method, a gene fusion mutation detection device, computer equipment and a computerstorage medium. The above-mentioned gene fusion mutation library construction method and the gene fusion mutation detection method and device are based on a DNA probe hybridization capture multi-geneRNA targeted sequencing technology, through a fusion gene capture probe hybridization capture target fusion gene, the gene fusion mutation library is constructed, the library can be used for high-throughput sequencing and bioinformatics analysis can identify core genes and partner genes there of constitute breakpoints. Furthermore, the present invention also designs a quantitative analysis methodof the fusion gene, a mutation ratio of the fusion gene can be obtained by calculation, accurate expression value of the fusion gene can be further obtained, and the quantitative analysis method of the fusion gene is pioneering and solves a quantitative analysis problem of using a NGS method to detect the fusion gene.
Owner:GUANGZHOU KINGMED DIAGNOSTICS GRP CO LTD
A markerless gene deletion attenuated mutant strain of Vibrio alginolyticus wild strain, related preparations and applications
The invention relates to an unmarked gene-deleted and attenuated mutant strain of a vibrio alginolyticus wild strain, which is the attenuated active vaccine of the vibrio alginolyticus wild strain and in which the sRNA molecular chaperone gene hfq of the vibrio alginolyticus wild strain is deleted. The vibrio alginolyticus wild strain is EPGS020401, and the collection number of the vibrio alginolyticus wild strain is CCTCC NO.AB 209306. The invention also provides the preparation of the unmarked gene-deleted and attenuated mutant strain of the vibrio alginolyticus wild strain and the use thereof in prevention and control of vibrio alginolyticus disease in cultured fish. The unmarked gene-deleted and attenuated mutant strain of the vibrio alginolyticus wild strain or the preparation thereof, which are disclosed by the invention, eliminates universal potential environment and product safety risks of the conventional attenuated vaccine and is a safe, effective and economic vaccine for preventing and controlling vibrio alginolyticus diseases in cultured fish.
Owner:EAST CHINA UNIV OF SCI & TECH
Preparation method of fusion gene positive control standard
ActiveCN109837275AAny convenient accessLow costMicrobiological testing/measurementDNA preparationPositive samplePositive control
The invention discloses a fusion gene positive control standard and a preparation method thereof. A fusion partner gene is amplified by a non-fusion gene positive sample, and the fusion partner gene nucleic acid fragment is ligated by asymmetric PCR combined with a self-annealing PCR method to obtain the corresponding fusion gene. Nested PCR and agarose gel electrophoresis are carried out for purification and recovery. After concentration determination, the corresponding fusion gene positive control standard is prepared. The fusion gene positive control standard and the preparation method thereof solve the problems that the fusion gene nucleic acid sequence positive control standard is difficult to obtain and the artificial synthesis cost is high. The fusion gene positive control standardhas application prospects in clinical fusion gene detection to assist diagnosis, treatment and prognosis. In addition, the synthetic fusion gene strictly controls the concentration of the two-side partner gene to 1:1, and can be used as an internal control for quantitative analysis such as gene copy number analysis. The fusion gene positive control product prepared by the preparation method has the advantages of simple preparation, flexibility, low cost and the like, and is of great significance in clinical detection, related quality inspection, and the like.
Owner:THE SECOND HOSPITAL OF DALIAN MEDICAL UNIV
Recombinant escherichia coli for producing pentamethylene diamine and application of recombinant escherichia coli
The invention discloses recombinant escherichia coli for producing pentamethylene diamine and application of the recombinant escherichia coli. A ribulose diphosphate carboxylase gene in rhodospirillum, a ribose phosphate kinase gene and a lysine decarboxylase gene in spinach are synthesized from the beginning, PCR copy an escherichia coli molecular chaperone gene to construct a plasmid pTrc99a-cadA-cbbM and a plasmid pCWJ-prkA-GroEL / GroES, the double-plasmid recombinant bacteria grow well in a specific culture medium, and glucose is efficiently utilized. According to the recombinant escherichia coli, inducers arabinose and IPTG are respectively added in different time periods, the heterologous gene expression is stable, the reaction system is simple, the condition is mild, the period is short, few byproducts are produced, the method is clean and pollution-free, the method is a simple, rapid and efficient production way, and the fermentation result shows that the yield of the recombinant escherichia coli KACCPG is obviously improved.
Owner:NANJING UNIV OF TECH
Construction and application of astaxanthin synthesis strain
ActiveCN112852694AImprove generation effectIncrease productionBacteriaMicroorganism based processesAstaxanthinPromoter
The invention discloses construction and an application of an astaxanthin synthesis strain. The invention provides a recombinant bacterium, which is prepared according to a method comprising the following steps of: expressing and synthesizing carotenoid related genes crtY gene, crtZ gene and crtW gene in escherichia coli, and regulating and controlling the expression of molecular chaperone genes groES and groEL in the escherichia coli to obtain the recombinant bacterium, according to the invention, the promoters with different intensities are utilized and gene copy is changed, so that coordinated expression among genes in a synthetic pathway of astaxanthin is realized, and the toxicity of metabolic intermediates is eliminated, so that the generation capacity of biosynthesis of astaxanthin is improved. In addition, stabilizing the expression of the exogenous gene and increasing the correct folding of the exogenous gene can also become an effective way for improving the biological activity of astaxanthin synthetic pathway enzyme and increasing the yield of astaxanthin.
Owner:TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
Process For Producing Polypeptide
A process for producing a target protein at low temperature, comprising inducing expression of not only a vector having introduced therein a gene coding for the target protein but also a vector having a chaperone gene introduced therein.
Owner:TAKARA HOLDINGS
Escherichia coli expression system containing manganese ion recombinant protein, and application method thereof
ActiveCN108998462ASimple separation and purification processLow total production costBacteriaHydrolasesEscherichia coliManganese
The invention discloses an Escherichia coli expression system containing a manganese ion recombinant protein, and an application method thereof. A recombinant expression plasmid in the Escherichia coli expression system is one or a combination of the following cases: (1) an Escherichia coli molecular chaperone gene and an manganese ion-containing enzyme protein gene; (2) an E. coli chaperone geneAn enzyme protein gene containing manganese ions, and a channel-associated protein gene for overexpressing or inhibiting manganese ions; and (3) the Escherichia coli molecular chaperone gene, the manganese ion-containing enzyme protein gene, and a concentration-associated protein gene for affecting intracellular manganese ions. The Escherichia coli expression system is constructed and optimized torealize the efficient, soluble and active expression of the manganese ion-containing enzyme protein; and compared with conventional methods, the method has the advantages of high efficiency, simple purification process, low cost, and facilitation of the industrial production and application of manganese ion-containing enzymes.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
FGFR gene fusion probes, detection method and kit
ActiveCN110218797AHigh sensitivityImprove featuresMicrobiological testing/measurementDNA/RNA fragmentationInteinExon
The invention relates to FGFR gene fusion probes, a detection method and a kit, and provides a probe combination for detecting human FGFR gene fusion. The probe combination comprises the following probes in a region of all exons and 1<st> intron, 15 intron and 17 intron of FGFR2 (NM_001144917), in a region of all exons, 17 intron, 18 intron and 3'UTR of FGFR3 (NM_000142) and in a region of 11 intron and 14 intron of an FGFR3 chaperone gene TACC3 (NM_006342). The invention also provides a kit for detecting FGFR gene fusion, in particular to a kit for detecting the FGFR gene fusion at a time and an automatic device used for analyzing human gene FGFR gene fusion. The FGFR gene fusion probes, the detection method and the kit are applicable to the detection of a wide range of samples.
Owner:上海思路迪医学检验所有限公司
Construction method of onion pseudomonas genetic engineering bacteria
InactiveCN101358195AHigh expressionHigh activityBacteriaMicroorganism based processesPseudomonasGenetically engineered
The present invention provides a method for constructing a genetically engineered bacterium from Pseudomonas cepacia, which replaces the lipase in the genome of Pseudomonas cepacia G63 and a chaperone gene with T7 RNA polymerase genes thereof, so that the T7 RNA polymerase genes are integrated into the genome of Pseudomonas cepacia and controlled by a lipase promoter. Afterwards, the Pseudomonas cepacia lipase and the chaperone gene thereof are cloned into expression plasmid containing T7 promoters and LacI repressor protein, and finally, the plasmid is delivered into the recombinant bacterium of Pseudomonas cepacia to obtain the super engineered bacterium of the Pseudomonas cepacia lipase. The method genetically modifies Pseudomonas cepacia G63, and the obtained super engineered bacterium has high expression and catalysis activity and good low-carbon alcohol tolerance and high-temperature resistance.
Owner:HUAZHONG UNIV OF SCI & TECH
Isolated nucleic acids encoding activated and suppressive forms of ATF6
InactiveUS6635751B1Good effectHigh activitySugar derivativesGenetic engineeringNucleotideReticulum cell
A factor capable of efficiently regulating expression of an endoplasmic chaperone gene, a nucleic acid encoding it, a complementary strand nucleic acid thereof, a method for regulating expression of an endoplasmic reticulum chaperone gene, and a method for expressing a foreign gene is provided. The endoplasmic reticulum stress transcription factor is capable of regulating transcription-inducing activity, wherein the transcription-inducing activity is exhibited by an element having the nucleotide sequence shown in SEQ ID NO:1 or an element having a nucleotide sequence resulting from substitution of 1 to 3 bases with other bases in the nucleotide sequence as shown in SEQ ID NO:1. A method for controlling expression of an endoplasmic reticulum chaperone, comprising expressing the factor; a method for expressing a foreign protein, comprising positively regulating expression of an endoplasmic reticulum chaperone gene by the method for controlling expression; and a nucleic acid encoding activated form of ATF6, activated form of CREB-RP, suppressive form of ATF6, or suppressive form of CREB-RP, or the complementary strand thereto are also provided. It is expected to be applied to treatment or prophylaxis of cancers, arteriosclerosis, cystic fibrosis, ischemic diseases, wounds or ulcers.
Owner:HSP RES INST
Recombinant plasmid for production of oxalate decarboxylase, system and method for expressing escherichia coli, and application
ActiveCN108977455AIncrease the concentration of manganese ionsOptimizing culture induction conditionsBacteriaPeptide/protein ingredientsBiotechnologySolubility
The invention discloses a recombinant plasmid for production of oxalate decarboxylase, a system and a method for expressing escherichia coli, and application. The recombinant plasmid for production ofoxalate decarboxylase comprises an oxalate decarboxylase gene, a molecular chaperone gene and a gene for regulating and controlling intracellular manganese ion concentration. The recombinant plasmidis guided into an escherichia coli expression bacterial strain and the escherichia coli with MntP gene deletion to respectively obtain oxalate decarboxylase with solubility expression and activity; the culture temperature and induction temperature of the recombinant bacterial strain are optimized; when the culture temperature is 37 DEG C, and the induction temperature is 25 DEG C, the expression amount of the oxalate decarboxylase is higher; by utilizing the two steps of organic solvent precipitation and ammonia sulfate precipitation to simply purify, the specific activity of the oxalate decarboxylase can reach 45.6U / mg, and the activity is higher than 95%. By adopting the technical scheme, the recombinant plasmid has the advantages that the production and purifying technologies are simple; the expression amount and specific activity of the oxalate decarboxylase are higher, the industrialized amplification is easy, the cost is low, and the recombinant plasmid is suitable for the industrialized production and application of the oxalate decarboxylase.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Method for simultaneously enhancing expression quantity and solubility of target protein in prokaryotic system
ActiveCN114480460AHigh expressionImprove solubilityPolypeptide with localisation/targeting motifMicroorganism based processesEscherichia coliProtein target
The invention discloses a method for simultaneously enhancing the expression quantity and solubility of a target protein in a prokaryotic system, and the method is used for guiding the expression of the target protein by using a novel fusion tag based on a Vip3 protein signal peptide sequence or a local sequence of the Vip3 protein signal peptide sequence. A fusion gene formed by the fusion tag and a coding gene of a target protein is co-expressed with an escherichia coli molecular chaperone Trigger factor gene or a homologous gene thereof in other organisms in the same prokaryotic expression system, so that the expression quantity and the solubility of the target protein can be enhanced. The importance of the invention lies in that a novel fusion tag is invented, the expression quantity of the target protein can be remarkably enhanced, and meanwhile, the tag can interact with a known specific molecular chaperone, so that the solubility of the target protein can be foreseeably enhanced on the premise of ensuring the expression quantity, one-by-one attempts on different molecular chaperones are avoided, and the target protein solubility is improved. The traditional scheme for enhancing the solubility of the unknown target protein can be determined, and the efficiency is greatly improved.
Owner:SHANXI AGRI UNIV
Method for preparing thanatin based on escherichia coli prokaryotic expression system
InactiveCN102943077AFast preparationEfficient manufacturingBacteriaMicroorganism based processesEscherichia coliCyanogen bromide
The invention relates to a method for preparing thanatin based on an escherichia coli prokaryotic expression system and belongs to the technical field of genetic engineering. A recombination gene comprises a fusion partner gene sequence, a methionine codon, a thanatin repetitive sequence and a stop codon, which are sequentially connected. The invention further provides a prokaryotic expression plasmid containing the recombination gene and escherichia coli containing the prokaryotic expression plasmid. The method for preparing the thanatin based on the escherichia coli prokaryotic expression system is characterized by comprising the following steps of (1) inducing escherichia coli to express recombination protein; (2) separating and purifying the recombination protein; (3) treating by cyanogen bromide to cut the recombination protein into a plurality of independent and complete thanatin; and (4) separating and purifying the thanatin. The innovation of the invention is as follows: the plurality of independent and complete thanatin is obtained in one step through splitting of the cyanogen bromide; and the expression quantity of the thanatin is high.
Owner:HANGZHOU SILKWORM TREASURE BIOLOGICAL TECH CO LTD
Activator of gene expression of molecular chaperone gene comprising eggshell membrane component and composition thereof
InactiveUS20150196606A1Reduce riskEasy to producePowder deliveryMammal material medical ingredientsEggshell membraneGene expression
Owner:THE UNIV OF TOKYO +1
Escherichia coli expression system for production of oxalate oxidase, production method of oxalate oxidase and application thereof
ActiveCN109022471AHigh specific vitalitySimple separation and purification processPeptide/protein ingredientsMicrobiological testing/measurementOxalate oxidaseEukaryotic plasmids
The invention discloses an Escherichia coli expression system for producing oxalate oxidase, a production method of oxalate oxidase and application thereof. The recombinant expression plasmid in the E. coli expression system comprises: an oxalate oxidase gene, a chaperone gene, and a gene that promotes to pump manganese ions into E. coli cell-associated protein. By screening and testing the genesof different manganese ion-related proteins and their copy number, the type of a promoter for controlling expression of the molecular chaperone and the type of a culture medium used in the productionprocess of oxalate oxidase, an optimized expression combination of oxalate oxidase is obtained, and the expression activity of oxalate oxidase in E. coli exceeds 80 U / mL. The specific activity of theoxalate oxidase preliminarily purified by a supernatant of a cell disrupted liquid is greater than 20 U / mg, and the molecular weight of the monomer is about 25 kDa. The technical scheme of the invention has the advantages of simple production and purification process, high expression level and specific activity, easy industrial amplification and low cost, and is advantageous for industrial production and application of such enzymes.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Method for improved high secretory production of proteins
The invention provides a method for improved high secretory production of proteins. The present invention addresses the problem of providing a production system which achieves, in a host cell such as yeast, high secretory production of proteins, particularly proteins having a complex structure such as an S-S bond and which is so safe as to obviate explosion-proof equipment and is suitable for industrial production. The present invention provides a transformed yeast obtained by introducing a chaperone gene and disrupting aox1 and / or protease genes, and a method for producing proteins using the transformed yeast.
Owner:DAIICHI SANKYO CO LTD
Method for preparing recombinant human nerve growth factor by using Escherichia coli expression system
ActiveCN102839182BHigh bioactivityEasy to prepareFermentationVector-based foreign material introductionEscherichia coliInclusion bodies
The invention discloses a method preparing a recombinant human nerve growth factor by using an Escherichia coli expression system. Beta-subunit genes of the recombinant human nerve growth factor comprise 6 histidine purification sites, an enterokinase cleavage site and a beta-subunit gene encoding mature peptide of modified human nerve growth factor and a molecular chaperone gene. The beta-subunit genes of the recombinant human nerve growth factor have high expression quantity in the Escherichia coli; generated inclusion body protein has high-efficiency renaturation under the assist of the molecular chaperone; and the recombinant human nerve growth factor obtained by nickel column affinity chromatography after the molecular chaperone is cleavaged accurately by the enterokinase, has a purity greater than 99% and biological activity greater than 500,000 AU / mg.
Owner:WUHAN HITECK BIOLOGICAL PHARMA
Folding of recombinant proteins via co-expression of archaeal chaperones
InactiveUS20120064560A1Easy to foldImprove the level ofSugar derivativesBacteriaBacteroidesProtein insertion
The present invention relates to recombinant protein production, and more specifically, to methods for recovery of properly folder bioactive proteins by expressing chaperone genes from extremophilic Archaea, during recombinant protein synthesis in a host cell thereby significantly improving recovery of properly folded bioactive proteins.
Owner:UNIV OF MARYLAND BALTIMORE
Dipel molecular companion gene, carrier containing same and bacterial strain
The present invention relates to a molecular chaperone gene p21zb cloned from Bacillus thuringiensis (Bt) and its carrier containing said carrier and strain. The transfer of said gene into Bt strain not only can obviously raise the yield of inserticidal crystal protein (ICPs) of wild type Bacillus thuringiensis, but also can excite high-level expression of insecticidal crystal protein silent gene of Bacillus thuringiensis. Said invention creates the recombinant carrier and engineering strain containing p21zb gene, and the ICPs expression quantity of said engineering strain is raised by above once as compared with that of initial strain. The tests show that said engineering strain possesses high toxicity for lepidopterous pests and good genetic stability, and has high-yield and high-toxicity property.
Owner:SUN YAT SEN UNIV
Construction and Application of Astaxanthin Synthetic Strain
ActiveCN112852694BImprove generation effectIncrease productionBacteriaMicroorganism based processesAstaxanthinPromoter
The invention discloses the construction of an astaxanthin synthetic strain and its application. The invention provides a recombinant bacterium, which is prepared according to a method comprising the following steps: expressing the synthetic carotenoid-related genes crtY gene, crtZ gene and crtW gene in Escherichia coli, and regulating the expression of molecular chaperone genes groES and groEL in the Escherichia coli , the obtained recombinant bacteria; the present invention realizes the coordinated expression of genes in the astaxanthin synthesis pathway by using promoters of different strengths and changing the gene copy, eliminates the toxicity of metabolic intermediates, thereby increasing the production capacity of astaxanthin biosynthesis. In addition, stabilizing the expression of exogenous genes and increasing the correct folding of exogenous genes can also be an effective way to improve the biological activity of enzymes in the astaxanthin synthesis pathway and increase the production of astaxanthin.
Owner:TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
Primers, probes and kits for detecting fusion mutations of human alk gene and their detection methods
ActiveCN109706232BSolve the problem of low coverageSensitive highMicrobiological testing/measurementDNA/RNA fragmentationMedicineExon
The invention discloses a primer, a probe, a kit and a detection method for detecting fusion mutations of human ALK gene, and the primer and probe include: designing on the broken exon sequences of six partner genes in the ALK fusion gene Multiple upstream primers, shared downstream primers and fluorescent detection probes designed on ALK exon 20; 6 partner gene break exon sequences in ALK fusion gene include: EML4‑exon2 / 6 / 13 / 14 / 17 / 20, TFG ‑exon5, KIF5B‑exon15 / 17 / 24, TPR‑exon15, HIPI‑exon21, SEC31A‑exon21; the kit can cover up to 28 ALK fusion forms, with high specificity and sensitivity, and can effectively avoid false positives. The established one-step reverse transcription detection method can simplify operation and avoid contamination.
Owner:合肥欧创基因生物科技有限公司
Escherichia coli expression system for producing oxalate oxidase, production method and application of oxalate oxidase
ActiveCN109022471BIncrease the concentration of manganese ionsHigh expressionPeptide/protein ingredientsMicrobiological testing/measurementOxalate oxidaseOxidative enzyme
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Molecular chaperone expression vector and strain for improving secretory expression of phytase in pichia pastoris
PendingCN113528565AOvercoming the insignificant increase in expressionOvercome the disadvantages of being less effective than a single molecular chaperoneFungiHydrolasesPhytaseSecretion expression
The invention relates to the field of gene engineering, in particular to a molecular chaperone expression vector and a strain for improving secretory expression of phytase in pichia pastoris. The expression vector comprises molecular chaperone genes BIP and PDI which are connected in series, or molecular chaperone genes VHB and PDI which are connected in series, or molecular chaperone genes HACI and PDI which are connected in series. A high-temperature-resistant phytase high-enzyme-activity expression strain V28-VTR-001 which is obtained through genetic engineering modification and screening and contains the expression vector is recombined pichia pastoris, and the total fermentation enzyme activity of 50 L of the recombined pichia pastoris reaches 37239 U / mL.
Owner:GUANGDONG VTR BIO TECH
Method for improved high-level secretory production of protein
InactiveUS20170211063A1Reduce the amount requiredImprove securityPeptidesOxidoreductasesBiotechnologyChaperonin
The object of the present invention is to provide a production system that is capable of high-level secretory production of a protein (and in particular, a protein with a complicated structure such as a structure with S—S bonds) in a host cell such as yeast and is suitable for industrial production with high safety that does not require explosion-proof facilities. The present invention provides a transformed yeast into which a chaperone gene has been introduced and in which the aox1 gene and / or the protease gene have been disrupted and a method for producing a protein involving the use of such transformed yeast.
Owner:DAIICHI SANKYO CO LTD
Mouse epigenetic gene knockout screening library and construction method thereof
ActiveCN112853502ASmall plasmidEasy to operateNucleotide librariesVector-based foreign material introductionEnzyme GeneArginine
The invention relates to a mouse epigenetic gene knockout screening library and a construction method thereof; the mouse epigenetic gene knockout screening library comprises SgRNA expression plasmids targeting epigenetic related genes; wherein The targeted epigenetic related genes comprise: a lysine acetyltransferase gene, a histone deacetylase gene, a histone lysine methyltransferase gene, a histone lysine demethylase gene, a protein arginine methyltransferase gene, a histone recognition and DNA methylation related gene, an RNA methylation related gene, a chromatin remodeling compound gene, andchromatin assembly and histone chaperone genes. The gene library constructed by the invention is a double-plasmid expression system, plasmids are relatively small, and construction is relatively easy to operate; meanwhile, the screening library only aims at epigenetic genes, so that the library is relatively small is size, and data analysis is relatively easy for single cell sequencing.
Owner:THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
A kind of genetically engineered bacteria expressing microbial cholesterol esterase and its construction method
The invention discloses a genetically engineered bacterium expressing microorganism cholesterol esterase and a construction method thereof, and belongs to the technical field of enzyme engineering. The method comprises the steps that Burkholderia cepacia. source cholesterol esterase and cholesterol esterase molecular chaperone genes are connected in series to achieve coexpression, wherein cholesterol esterase genes and the molecular chaperone genes are connected through a SD sequence. According to the method, the Burkholderia cepacia. source cholesterol esterase is successively expressed, and active expression and efficient expression of the Burkholderia cepacia. source cholesterol esterase are achieved. The cholesterol esterase activity in a crude enzyme solution fermented by the constructed genetically engineered bacterium can reach 640 U / L.
Owner:JIANGNAN UNIV
Escherichia coli expression system and application method of recombinant protein containing manganese ions
ActiveCN108998462BSimple production processOptimized to increase the concentration of manganese ionsBacteriaHydrolasesEscherichia coliIonic Channels
The invention discloses an Escherichia coli expression system of recombinant protein containing manganese ions and an application method thereof. The recombinant expression plasmid in the Escherichia coli expression system is one of the following situations or a combination thereof: (1) at least comprising an Escherichia coli molecular chaperone gene and an enzyme protein gene containing manganese ions; (2) comprising an Escherichia coli molecular chaperone gene , enzyme protein genes containing manganese ions, and overexpression or inhibition of manganese ion channel-related protein genes; (3) including Escherichia coli molecular chaperone genes, enzyme protein genes containing manganese ions, and protein genes affecting intracellular manganese ion concentration . Through the construction and optimization of the Escherichia coli expression system, efficient soluble and active expression of the manganese ion-containing enzyme protein can be realized. Compared with the traditional method, the efficiency is high, the purification process is simple, and the cost is low, which is beneficial to the production of the manganese ion-containing enzyme. Industrial production and application.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Light-emitting reporting system and application thereof to antibiotic discovering and screening
ActiveCN110241129ARapid identificationHigh-throughput identificationMicrobiological testing/measurementVector-based foreign material introductionScreening techniquesHigh flux
The invention discloses a light-emitting reporting system. The light-emitting reporting system is constructed from a promoter fragment of molecular chaperone genes clpX, groE, dnaJ and dnaK, and a luxABCDE gene fragment connected with molecular chaperone gene promoters. The invention further discloses an application of the light-emitting reporting system to antibiotic recovering and screening. Compared with an existing antibiotic screening technique, the light-emitting reporting system disclosed by the invention has the advantages that the existence of antibiotics in an solution can be identified in a quick and high-flux manner, the discovering speed of new antibiotics can be increased, quick discovering and screening of the antibiotics can be realized, and the light-emitting reporting system has great application prospects.
Owner:SHANDONG UNIV
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