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87 results about "Polymerase Gene" patented technology
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Polymerase Genes encode enzymes (Polymerases) that catalyze polymerization reactions, especially of nucleotides to polynucleotides. (NCI)
Method for specific integration of t7 rna polymerase gene in the chromosome of corynebacterial and the resultant corynebacteria-t7 promoter based shuttle vector system
The present invention relates to method for obtaining optimum expressed proteins in a transformed gram positive bacteria by specific integration of T7 RNA polymerase gene into the chromosome of a gram positive bacteria exhibiting resistance to aminoglycosides, said method comprising:- digesting an E. coli plasmid with a restriction enzyme- digesting the genomic DNA of said gram positive bacteria- ligating the said digested plasmid to the digested genomic DNA of said gram positive bacteria- transforming the said gram positive bacteria protoplasts with the said ligation mixture of step 2 to yield transformed gram positive bacteria (transformants),- screening the said transformants for kanamycin resistance and aminoglycoside sensitivity to ensure that the targetting of the said plasmid vector into the chromosome of the said gram positive bacteria is successful- cloning of the desired gene in the said vector—culturing the transformant in a suitable culture medium- isolating the expressed proteins from the culture medium
Owner:INDIAN INST OF TECH
Double-control double-regulation prokaryotic expression vector system and construction method and application thereof
InactiveCN102174554AHigh broad spectrumStrong broad spectrumVector-based foreign material introductionVector systemProkaryotic expression
The invention belongs to the field of molecular biological gene engineering, and particularly relates to a double-control double-regulation prokaryotic expression vector system and a construction method and application thereof. Two expression vectors finish expression of a target gene in the vector system, wherein one expression vector is a main expression vector, and the target gene is cloned on the expression vector; the other expression vector is an auxiliary expression vector, and the auxiliary expression vector controls a switch for regulating the promoter of the main expression vector; the main expression vector contains an SP6 promoter and a lactose regulating gene; the auxiliary expression vector contains an arab promoter, an araC regulating gene and an SP6RNA polymerase gene; and the target gene expressed by the main expression vector and the auxiliary expression vector are controlled and / or regulated by an inducer.
Owner:INNER MONGOLIA UNIV FOR THE NATITIES
Poly(ADP-ribose) polymerase-gene
The invention relates to poly(ADP-ribose)polymerase (PARP) homologs which have an amino acid sequence which hasa) a functional NAD+ binding domain andb) no zinc finger sequence motif of the general formulaCX2CXmHX2C in which m is an integral value from 28 or 30, and the X radicals are, independently of one another, any amino acid;and the functional equivalents thereof; nucleic acids coding therefor; antibodies with specificity for the novel protein; pharmaceutical and gene therapy compositions which comprise products according to the invention; methods for the analytical determination of the proteins and nucleic acids according to the invention; methods for identifying effectors or binding partners of the proteins according to the invention; novel PARP effectors; and methods for determining the activity of such effectors.
Owner:BASF AG +1
Recombinant strain for producing 3-hydracrylic acid homopolymer and/or 3-hydracrylic acid copolymer and application thereof
ActiveCN102174542AImprove conversion efficiencySimple production processBacteriaMicroorganism based processes3-Hydroxypropionic acidPolymerase L
The invention discloses a recombinant strain for producing a 3-hydracrylic acid homopolymer and / or a 3-hydracrylic acid copolymer and an application thereof. The construction method of the recombinant strain comprises the following steps: leading 1,3-Propanediol dehydrogenase coded genes, aldehyde dehydrogenase coded genes, 3-hydracrylic acid coenzyme A ligase coded genes and PHA (Polyhydroxyalkanoates) polymerase coded genes into a starting strain to obtain the recombinant strain. The experiments in the invention prove that the engineering bacteria can efficiently express 3-hydracrylic acid coenzyme A ligase coded genes and PHA polymerase coded genes and enable the 3-hydracrylic acid to be finally polymerized into 3-hydracrylic acid homopolymer (P(3HP)) from the 3-hydracrylic acid coenzyme A. Minitype fermentation tank experiments show that the engineering bacteria provided by the invention can have a maximum P (3HP) output of 8.9g / L after being fermented in a 6L fermentation tank and the P (3HP) can account for a maximum 91.5% of cell dry weight. In addition, the recombinant strain provided by the invention has the advantages of simple production process, low costs and broad application prospects.
Owner:TSINGHUA UNIV
Recombinant Taq DNA polymerase and preparation method thereof
ActiveCN103602643AImprove expression efficiencyImprove purification efficiencyTransferasesMicroorganism based processesActive proteinGenetic engineering
The invention relates to the technical field of genetic engineering, and particularly relates to a method for preparing a gene recombinant expression enzyme of Thermusaquaticus TaqDNA polymerase. The nucleotide sequence of a gene and the amino acid sequence of an encoded protein are respectively shown as SEQIDNO:1 and SEQIDNO:2. According to the invention, the method comprises the following steps: cloning a gene sequence by using a gene cloning technique, and building a prokaryotic expression vector; by using a recombinant strain of a built TaqDNA polymerase gene, carrying out suspension by using an LB culture solution; after carrying out culturing for 4.5 hours at a temperature of 37 DEG C, adding IPTG (isopropyl-beta-d-thiogalactoside) with a final concentration of 0.5 mM into the obtained substance to induce for 16-20 hours; after thalli are collected in a centrifugal mode, dissociating the thalli by using 2-4 mg / ml lysozyme, so that protein fluid is obtained; and filtering the protein fluid by using a chromatographic column and a dialysis bag so as to obtain high-purity active protein TaqDNA polymerase. Compared with the prior art, stability of a production process is good, activity of obtained TaqDNA polymerase is high, the yield and purity of products are effectively ensured, and amplification effect when being applied to a gene PCR (polymerase chain reaction) is good.
Owner:哲尔基因科技(上海)有限公司
ST cell lines for stably expressing T7 RNA polyase, constructing process and applications thereof
InactiveCN101285054AEnzymesVector-based foreign material introductionMicroorganism preservationFluorescence
The invention discloses a cell line ST / T7 capable of stably expressing T7RNA polymerase, the microorganism preservation number of which is: CGMCC No.24444. In the invention, a T7RNA polymerase gene is inserted into a eukaryotic expression vector pIRES2-EGFP to get the pIRES2-EGFP-T7 to transfect ST cells to obtain the cell line ST / T7 capable of stably expressing T7RNA polymerase. Through RT-PCR detection, expression plasmid containing a red fluorescent protein gene controlled by T7 promotors can be used to transfect the cell line so as to observe the expression of red fluorescent protein in a transcription product amplified form a ST / T7 cell to the T7RNA polymerase. The cell line ST / T7 can provide reverse T7RNA polymerase for use in reverse genetic manipulation of RNA-virus such as hog cholera virus, etc., and can be used as transcription and expression systems in vitro for research on genic structures and functions.
Owner:HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
Genetic engineering bacterium for L-theanine production and fermentation method thereof
ActiveCN109370966AClear genetic backgroundObvious cost advantageBacteriaTransferasesEscherichia coliT7 phage
The invention provides a genetic engineering bacterium for L-theanine production and a fermentation method thereof. A construction method of the genetic engineering bacterium comprises the steps of carrying out single copying on an ribonucleic acid (RNA) polymerase gene T7RNAP derived from T7 phage on an Escherichia coli W3110 genome, wherein the gene is controlled by a xylose promoter; carrying out dual copying on a gamma-glutamyl methylamine synthetase gene gmas derived from methylovorusmays, wherein the gene is controlled by a T7 promoter; knocking out a xylose operon repressor protein genexylR; and knocking out a succinyl CoA synthetase gene sucCD. The genetic engineering bacterium has the beneficial effects that the yield of L-theanine fermented by using the genetic engineering bacterium can be up to 40g / L and the sugar acid conversion rate can be up to 25%; and the L-theanine production method provided by the invention has the advantages of being low in raw material price, shortin cycle, simple and convenient to operate, green and friendly to environment and high in yield and has good industrial application value.
Owner:TIANJIN UNIV OF SCI & TECH +1
Recombinant bacteria for producing salidroside and analogue thereof, and construction method and use
InactiveCN107460203AResolve source issuesReduce manufacturing costBacteriaMicroorganism based processesSalidrosideMicrobiology
The invention discloses recombinant bacteria for producing salidroside and analogue thereof, and a construction method and use. The construction method comprises the steps of synthesizing keto-decarboxylase gene synkdc and glycosyl transferase gene synyjic, and connecting to a vector pRSFDuet-1 to construct a recombinant vector pSynkdc1-yjic; enabling T7RNA polymerase gene to be free or integrated to SyBE-002447 base bacterial strain, and introducing pSynkdc1-yjic to obtain SyBE-218011 and SyBE-218013 bacterial stains; and directly integrating synkdc and synyjic to SyBE-002447 chromosome, and removing feaB and ushA in a knockout manner to obtain a SyBE-218015 bacterial strain. The bacterial strain disclosed in the invention can be used for producing salidroside and analogue thereof through fermentation, so that the problem existing in source of the salidroside and analogue thereof can be solved, and cost can be lowered.
Owner:TIANJIN UNIV
siRNA capable of inhibiting foot and mouth disease virus replication and infection and its preparation method
InactiveCN1757725ATranscription assuranceEfficient transcriptionViruses/bacteriophagesAntibody medical ingredientsViral replicationDomestic animal
A kind of siRNAs able to suppress the copy and infection of foot-and-mouth disease virus in cell and animal individual and its preparing process and application are disclosed. The 1D protein gene and 3D polymerase gene are chosen as the interference targets from the genome of foot-and-mouth disease virus. The human recombinant adenovirus Ad5 is designed and used as the carrier of three siRNAs. The target of 21VPI siRNA and 63VPI siRNA is said 1D gene. The target of 56POL siRNA is 3D gene.
Owner:FUDAN UNIV
Iterative gene circuit based on vibrio fischeri quorum sensing system and T7 expression system, and application of iterative gene circuit
The invention discloses an iterative gene circuit based on a vibrio fischeri quorum sensing system and a T7 expression system. The iterative gene circuit comprises a signal molecular protein gene luxIof vibrio fischeri, a receptor protein gene luxR, a promoter sequence PluxI(1) for sensing a signal molecular and receptor protein complex, a T7 RNA polymerase gene T7 RNApoly and a green fluorescentprotein egfp for representing the circuit intensity. The gene circuit does not rely on an inducer and can utilize the T7 expression system to spontaneously and forcefully initiate target gene expression in any host bacterium; and gene elements are constructed on different expression vectors, the iterative gene circuit is constructed in engineering bacteria, the T7 expression system is utilized toamplify signals of the vibrio fischeri quorum sensing system, and the iterative gene circuit has the function of utilizing the T7 expression system to spontaneously and forcefully initiate target gene expression, and has a lower leakage ratio and higher initiating strength.
Owner:NANJING AGRICULTURAL UNIVERSITY
PNA Probes, Kits, and Methods for Detecting Lamivudine-Resistant Hepatitis B Viruses
InactiveUS20080233557A1High sensitivityStrong specificitySugar derivativesMicrobiological testing/measurementHbv dna polymeraseLamivudine resistance
Disclosed are peptide nucleic acid (PNA) probes to detect lamivudine resistant mutants of hepatitis B virus (HBV), which causes acute and chronic hepatitis, kits for detecting lamivudine resistant HBV comprising the PNA probes, and methods for detecting lamivudine resistant HBV by using the kits. They can accurately detect mutations of rtL180M, rtM204V, rtM204I and rtV2071 within B and C domains of HBV DNA polymerase gene, the main cause of lamivudine resistance, as well as mixed mutants of more than one mutant. They can detect lamivudine resistant HBV with high specificity and sensitivity.
Owner:PANAGENE INC
Detection method of hepatitis B virus drug-resistant mutation
InactiveCN106367528AImprove detection efficiencyDetection time is shortMicrobiological testing/measurementFluorescenceHBV polymerase
The invention discloses a detection method of hepatitis B virus drug-resistant mutation, and relates to hepatitis B viruses. The detection method comprises the following steps: 1) designing a PCR amplification primer in a relatively conservative area on an HBV (hepatitis B virus) polymerase gene sequence; 2) designing a fluorescent probe which can cover HBV drug-resistant mutation sites; 3) identifying characteristic melting-point temperature values of the fluorescent probe and the PCR amplification product after hybridization by virtue of a melting curve analysis technology; and 4) judging whether the drug-resistant mutation occurs in the prepared HBV polymerase gene sequence, and identifying the type of the HBV drug-resistant mutation. The detection method is comprehensive in the coverage of drug-resistant mutation sites, high in detection efficiency and short in consumed detection time; the detection adopts homogeneous detection and closed-tube operation; the detection method is simple to operate and high in detection throughput; and the detection method is high in detection specificity and is capable of judging and reading results easily.
Owner:XIAMEN UNIV +1
Paeonia ostii reference gene under drought stress, and special primer and application thereof
ActiveCN111733168AImprove stabilityImprove reliabilityMicrobiological testing/measurementOxidoreductasesBiotechnologyInitiation factor
The invention relates to a paeonia ostii reference gene under drought stress, and a special primer and application thereof. The reference gene is a TATA box binding protein TBP gene, an actin ACT1 gene, an actin ACT2 gene, a glyceraldehyde-3-phosphate dehydrogenase GAPDH gene, an eukaryote translation initiation factor eIF1 gene, a eukaryote translation initiation factor eIF2 gene, a tubulin alpha-TUB gene, a tubulin beta-TUB gene, an RNA polymerase II RNA Pol II gene or an RNA polymerase II transcription factor RP II gene. The nucleotide sequence of the gene is disclosed as the sequence table. The invention aims to provide a gene which can be stably expressed in the paeonia ostii gene expression profile under drought stress, and the gene is used as a paeonia ostii drought stress referencegene.
Owner:YANGZHOU UNIV
Recombinant expression plasmid based on T7 promoter, and transformant and application thereof
PendingCN108456688APrevent growth inhibitionHigh copy numberVectorsBacteriaBiotechnologyFermentation
The invention relates to a recombinant expression plasmid, especially to a recombinant expression plasmid based on a T7 promoter. The invention also relates to a transformant containing the recombinant expression plasmid, and application of the transformant, particularly application of the transformant to fermentation production of lysine decarboxylase and the like and production of 1,5-pentamethylene diamine. The recombination expression plasmid comprises a plasmid skeleton; a replicon; a target gene and the T7 promoter controlling the expression of the target gene; and a T7 RNA polymerase gene and a sugar-induced stringent promoter controlling the expression of the T7 RNA polymerase gene, e.g., an Arabinose promoter. The transformant contains the recombinant expression plasmid as described above. The invention provides a method for fermentation production of polypeptide. The method comprises a step of culturing any transformant as described in the specification. According to the invention, cost for current induced recombinant expression inducers is lowered.
Owner:CATHAY R&D CENT CO LTD +1
Universal primer sequence for detecting human coronavirus infection and detection method
InactiveCN104498636AMicrobiological testing/measurementAgainst vector-borne diseasesNucleotideNucleic acid
The invention designs a pair of degenerate primers hCoV-For and hCoV-Rev applied to universal qualitative detection of human coronavirus in a genomic polymerase gene coding region (1b region) by virtue of comparison analysis of six human coronavirus complete genomic nucleotide sequences logged in GenBank. An upstream primer sequence hCoV-For is as follows: ATGGGWTGGGAYTAYCCHAARTGTG; and downstream primer sequence hCoV-Rev is as follows: TGYTGKGARCARAAYTCRTGWGG; the size of an amplification target fragment is 605bp; RNA of positive specimens of HCoV-229E, HCoV-OC43, HCoC-NL63 and HCoV-HKU1 are respectively extracted for testing the universal primer by One-step RT-PCR. Results show that four human coronavirus RNAs are used as templates, and hCoV-For and hCoV-Rev are used as the universal primers for carrying out One-step RT-PCR reaction, a single target strip with the size about 606p can be amplified; by carrying out sequencing validation of PCR products, results show that the amplification sequence is right without cross reaction. The universal primer sequence can be applied to quick and qualitative detection of nucleic acid of human coronavirus infection.
Owner:江苏国际旅行卫生保健中心无锡分中心
Method for producing short-and-medium-chain-length polyhydroxyalkanoate (PHA) and functional derivatives thereof
ActiveCN111235173AControllable ratioReduce energy consumptionBacteriaMicroorganism based processesLong chain fatty acidBinding site
The invention discloses a construction method of a recombinant bacterium for producing short-and-medium-chain-length PHA and functional derivatives thereof. The method comprises the following steps of: introducing a coding gene of specific PHA polymerase, a coding gene of a key protein of a PHA synthesis pathway, a promoter or a mutated promoter, related genes capable of enhancing carbon source utilization capacity of medium-chain and long-chain fatty acids, the ribosome binding site RCJ and the ribosome binding site RD into a starting strain from which an endogenous PHA polymerase gene is knocked out; and adjusting the proportions of all monomers in the short-and-medium-chain-length PHA and the functional derivatives thereof in the recombinant bacteria so as to realize controllable production of the short-and-medium-chain-length PHA and the functional derivatives thereof.
Owner:TSINGHUA UNIV
Method for controlling CSBV (Chinese bee Sacbrood Virus) by utilizing RNA interference technology
ActiveCN103751807AInhibition of replicationLow costGenetic material ingredientsAntiviralsEscherichia coliChinese bee
The invention relates to a method for controlling CSBV (Chinese Bee Sacbrood Virus) by utilizing an RNA interference technology. The method comprises the following steps: synthesizing dsRdRp through a bacteria prokaryotic induction expression method by utilizing an RNA-dependent RNA polymerase (RdRp) gene which is specific to the CSBV and related to virus replication, and adding double-strand RNA into feed for feeding larva of 1-2 days. The double-strand RNA is expressed by Escherichia coli induction, is low in cost and has a good curing effect for controlling the CSBV. The feeding concentration of dsRdRp is 0.5 microgram / ml, and the pupation rate and eclosion rate of the larva are respectively 70 percent and 55 percent and are obviously improved compared with the pupation rate of 10 percent and the eclosion rate of 0 percent in the larva fed of dsGFP.
Owner:FUJIAN AGRI & FORESTRY UNIV
Gamma-PGA polymerase gene recombinant strain as well as construction method and application thereof
ActiveCN112175982AIncrease productionRegulating Molecular WeightCosmetic preparationsBacteriaBiotechnologyBacillus licheniformis
The invention discloses a gamma-PGA polymerase gene recombination strain as well as a construction method and application thereof, relates to a production process for synthesizing gamma-polyglutamic acid from a sugar raw material through one-step fermentation, and belongs to the field of synthetic biology and fermentation engineering. According to the invention, a gene cluster capBCA of gamma-polyglutamic acid synthetase of naturally produced bacillus licheniformis is cloned into corynebacterium glutamicum F343 with high yield of glutamic acid for exogenous expression, and on the basis, the molecular weight of gamma-PGA produced by recombinant strains Cg 1 / 10BCA, Cg B1 / 10CA and Cg BC1 / 10A obtained by regulating and controlling the expression level of each gene of a synthetase gene clusterby utilizing RBS regulating and controlling elements with different strengths is 295.47-28018 kDa. According to the method, an exogenous synthesis path of polyglutamic acid is successfully constructed, the purpose of reasonably controlling the molecular weight of gamma-PGA is achieved, raw materials and process control cost are saved, economic benefits are improved, the application range of polyglutamic acid is expanded, and the method has very good industrial application value and prospect.
Owner:JIANGNAN UNIV
Specific primer, probe and quick detection kit for detecting macrobrachium rosenbergii flavivirus-1
ActiveCN110846441ASkip the electrophoresis stepEasy to operateMicrobiological testing/measurementMicroorganism based processesFluoProbesNucleotide
A primer probe mixed solution for detecting macrobrachium rosenbergii flavivirus-1 comprises a primer set A and a Taqman fluorescent probe B. The 5' end of the Taqman fluorescent probe B is marked with a fluorescent reporting group, and the 3' end of the Taqman fluorescent probe B is marked with a fluorescent quenching group. The primer set A comprises a primer MrFV-q150F and a primer MrFV-q150R.The nucleotide sequences of the primer MrFV-q150F and the primer MrFV-q150R are respectively represented by SEQ ID N0:1 and SEQ ID N0:2. The nucleotide sequence of the Taqman fluorescent probe B is represented by SEQ ID N0:3. A fluorescent quantitative detection kit for the macrobrachium rosenbergii flavivirus-1 comprises the mixed solution, an RT-qPCR reaction solution, a positive quality controland a negative quality control. The RT-qPCR reaction solution is an RT-qPCR reaction solution applied to the probe method and the fluorescent quantitation one-step method, the negative quality control is sterile normal saline, and the positive quality control is a vector containing polymerase genes of the macrobrachium rosenbergii flavivirus-1. The kit is used for detecting the macrobrachium rosenbergii flavivirus-1, and the speed and specificity are high.
Owner:ZHEJIANG INST OF FRESH WATER FISHERIES
DNA polymerase coding DNA, enzyme coded thereby, and application and preparation method of DNA polymerase
The invention belongs to the field of gene engineering, and relates to a DNA polymerase gene separated from Hyperthermophilic Archaeon, a plasmid thereof, a DNA polymerase coded thereby, a preparation method of the DNA polymerase, and an application of the DNA polymerase, and concretely relates to DNA polymerase coding DNA separated from Pyrococcus yayanosii, and an application and a preparation method thereof. The sequence of the DNA polymerase coding DNA is one of a nucleotide sequence represented by SEQ ID NO.1in a sequence table, and a nucleotide sequence obtained by deleting, inserting and substituting one or more nucleotides into the sequence represented by SEQ ID NO.1. The encoded DNA polymerase has the advantages of super strong high-temperature resistance, high fidelity performance, excellent anti-inhibitor ability, and realization of a direct PCR reaction by using crude samples.
Owner:SNOVA BIOTECH
Biochip for streptococcus pneumoniae serotype detection, and its detecting method and kit
InactiveCN1710108AImprove accuracyGood repeatabilityMicrobiological testing/measurementBiochipStreptococcus constellatus
The invention relates to one kind of gene chip used in the pneumonia streptococcus blood serum examination and its examination method and the reagent box used thereof. The gene chip includes the solid phase carrier and the oligonucleotide probe fixed on this carrier. The probe includes the DNA fragment selected from the streptococcus pneumoniae 16s rDNA and the DNA fragment selected from the polyase gene of streptococcus pneumoniae. Use designed primer to enlarge and mark the gene group of the sample to be detected, then to hybrid the gene group using the biological chip, the different type of serum of the streptococcus pneumoniae can be detected.
Owner:TIANJIN BIOCHIP TECH CO LTD
Code error-prone DNA (Deoxyribonucleic Acid) polymerase and preparation method thereof
ActiveCN108949719AIncrease vitalityError-prone effectTransferasesVector-based foreign material introductionPolymerase LThiogalactosides
The invention belongs to the technical field of genetic engineering and in particular relates to code error-prone DNA (Deoxyribonucleic Acid) polymerase and a preparation method thereof. The preparation method of the code error-prone DNA polymerase comprises the following steps: (1) extracting rhodococcus genome DNA; sequencing and comparing to obtain a code error-prone DNA polymerase gene sequence SEQ ID NO: 1; (2) cloning a nucleotide sequence shown as the SEQ ID NO: 1 so as to obtain a nucleotide segment; (3) connecting the nucleotide segment with a plasmid pET32a, so as to construct a recombinant plasmid; (4) transforming the recombinant plasmid to an escherichia coli competent cell E.coli Bl21, so as to obtain an expression strain; (5) culturing and expressing the strain by utilizingan LB liquid culture medium, and adding IPTG (isopropyl-beta-d-thiogalactoside) with the final concentration of 0.5mM to induce for 16 to 20h; and cracking thalli and purifying to obtain the recombinant error-prone DNA polymerase, wherein an amino acid sequence of the recombinant error-prone DNA polymerase is shown as the SEQ ID NO: 1. By adopting the preparation method, the error-prone DNA polymerase expression and purification efficiency is improved, and the stable and high-activity error-prone DNA polymerase is obtained; and the error-prone effect is improved.
Owner:SHANXI MEDICAL UNIV +1
Small interfering ribose nucleic acid (siRNA) targeting molecule and application thereof
ActiveCN101979556AInhibition of translationInhibition of replicationGenetic material ingredientsDigestive systemSense strandHepatitis B virus
The invention relates to a small interfering ribose nucleic acid (siRNA) targeting molecule and application thereof. The positive-sense strand of the siRNA targeting molecule is shown as SEQ ID NO:3, and the antisense strand of the siRNA targeting molecule is shown as SEQ ID NO:4, wherein the antisense strand is specifically combined with messenger ribose nucleic acid (mRNA) of a hepatitis B virus (HBV) polymerase gene to degrade the mRNA so as to interfere a post-transcriptional translation process and fulfill the aim of resisting HBV.
Owner:BIOMICS BIOTECH +1
Method and kit for detecting genotypes of herpes simplex virus
InactiveCN102071262AReliable resultsAvoid misdiagnosisMicrobiological testing/measurementMicroorganism based processesFluoProbesCycle threshold
The invention discloses a method and a kit for detecting genotypes of a herpes simplex virus (HSV). Primer probes designed on the basis of HSV DNA polymerase gene are adopted, type 1 and type 2 of the HSV are detected simultaneously in a single tube by TaqMan probe dual-wavelength fluorescent PCR technology, and the existence and the type of the HSV in a sample are judged through fluorescent signals and circulating threshold values of amplification templates with different wavelengths. In the kit, a manually constructed internal reference system is used for avoiding false negative of a detection result. The kit comprises nucleic acid extracting solution, PCR buffer solution, Taq enzyme, an internal reference, negative control and positive control; and two steps of sample treatment and amplification detection are adopted. The kit is easy and convenient to operate and high in sensitivity, can avoid false positive caused by PCR product nucleic acid pollution, can solve the problem of false negative caused by a PCR inhibitor in the sample, and can be widely applied to quick detection of the genotypes of the pathogen clinically.
Owner:SHANGHAI XINGYAO MED TECH DEV CO LTD +1
Detection primer group, detection method and kit for loop-mediated isothermal amplification of GI type Norovirus
ActiveCN103642943AStrong specificityHigh sensitivityMicrobiological testing/measurementDNA/RNA fragmentationWater bathsFluorescence
The invention discloses a detection primer group, a detection method and a kit for loop-mediated isothermal amplification of GI type Norovirus, wherein the detection primer group comprises a forward outer primer F3, a backward outer primer B3, a forward inner primer FIP and a backward inner primer BIP, which are shown in SEQ ID NO. 1, 2, 3 and 4, respectively. According to the invention, the LAMP detection method of the GI type Norovirus is established; the detection method is characterized in that a set of specific primers (four primers), including two specific inner primers and two specific outer primers, is designed and screened for the RNA (Ribose Nucleic Acid) polymerase gene of the GI type Norovirus. The detection method provided by the invention employs the LAMP technology and is high in specificity, and has higher sensitivity than the RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection method; however, the expensive PCR instrument is not needed and only a common water bath is needed; in addition, the results can be observed by using a fluorescent dye in stead of a gel electrophoresis method; therefore, the detection method is simple and quick, and can be applicable to detecting the GI type Norovirus, and in particular to the basement layer sites.
Owner:GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY +1
Method and kit for detecting nucleic acid with herpes simplex virus
InactiveCN102071261AAvoid misdiagnosisWill not interfere with each otherMicrobiological testing/measurementMicroorganism based processesPositive controlFluorescence
The invention discloses a method and a kit for identifying a herpes simplex virus (HSV) by utilizing fluorescent PCR. Primer probes designed on the basis of HSV DNA polymerase gene and a manually constructed internal reference system are adopted, the herpes simplex virus and internal reference DNA are detected in a single tube by TaqMan probe dual-wavelength fluorescent PCR technology, and the existence of the HSV in a sample is judged through fluorescent signal intensity and a circulating threshold value of an amplification template. The kit comprises nucleic acid extracting solution, PCR buffer solution, Taq enzyme, an internal reference, negative control and positive control; and two steps of sample treatment and amplification detection are adopted. The kit is easy and convenient to operate and high in sensitivity, can avoid false positive caused by PCR product nucleic acid pollution, can solve the problem of false negative caused by a PCR inhibitor in the sample, and can be widely applied to quick detection of the pathogen clinically.
Owner:SHANGHAI XINGYAO MED TECH DEV CO LTD +1
Hepatitis B virus multi-locus genotypic resistance mutation detection method
InactiveCN101698891AEliminate the effects ofHigh sensitivityMicrobiological testing/measurementMicroorganism based processesViral Reverse TranscriptionMutation detection
The invention discloses a hepatitis B virus multi-locus genotypic resistance mutation detection method. A single tube nest type polymerase gene amplification method for detecting reverse transcriptase full gene sequence of virus includes that nest type PCR amplification is carried out in a single tube for twice, products of a first group PCR reaction are all used as templates of a second group PCR reaction by adjusting concentration of internal and external primers, and finally nest type PCR reaction is completed in the same single tube. The invention ensures specificity and economy of detection while obviously improving detection sensibility and has the detection range over a hundred million difference viral load.
Owner:THE FIFTH MEDICAL CENT OF CHINESE PLA GENERAL HOSPITAL
Strain capable of improving performance of recombinant protein under aerobic condition
The invention provides a strain capable of improving the performance of recombinant protein under an aerobic condition, and belongs to escherichia coli. The strain comprises at least one non-endogenous promoter which is operably connected to a glpF gene, a gldA gene, a dhaKLM gene operator sequence and an araE gene on the chromosome of the strain, an araBAD promoter which is operably connected to a T7RNA polymerase gene on the chromosome, and a T7 promoter which is operably connected to a recombinant protein gene. Furthermore, the strain is lack of an araABD gene operator sequence and an araFGH gene operator sequence.
Owner:FENG CHIA UNIVERSITY
Cell line capable of realizing induced rescue of rhabdovirus as well construction method and application of cell line
InactiveCN107557340AImprove rescue efficiencyEasy to operateStable introduction of DNAMicroorganism based processesVirusGenome
The invention provides a cell line capable of realizing induced rescue of rhabdovirus as well a construction method and an application of the cell line. The cell line is a BHK(TetOn) cell line for performing lentivirus induced expression on T7pol / P / N. The rhabdovirus construction method comprises the following steps: transfecting dual plasmids, namely a virus polymerase gene plasmid pL and a rhabdovirus core framework plasmid pCore, into the BHK(T7pol / P / N-TetOn) cell line according to a specific ratio, thereby rapidly obtaining a recombinant rhabdovirus. A vector for performing induced expression on T7pol / P / N is integrated into a BHK(TetOn) cell genome through pGag / Pol, pRSV-Rev and pMD2.G lentivirus plasmid system. According to the cell line disclosed by the invention, the rhabdovirus assembly can be efficiently subjected to rapid rescue, and screening and identification can be completed. Compared with the traditional multi-plasmid rescue system, the plasmid system in the invention has the advantage that the rescue efficiency of the recombinant rhabdovirus is greatly improved.
Owner:南京普菲科医药科技有限公司
A kind of genetically engineered bacteria for l-theanine production and fermentation method thereof
ActiveCN109370966BClear genetic backgroundObvious cost advantageBacteriaTransferasesEscherichia coliEnzyme Gene
Owner:TIANJIN UNIV OF SCI & TECH +1
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