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41 results about "Polyadenylic acid" patented technology
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Multivalent immunostimulation nanoformula, preparation method and application thereof
InactiveCN103007294AIncrease uptakeVitality has no effectPowder deliveryOrganic active ingredientsPolyadenylic acidAqueous solution
The invention provides a preparation method of a multivalent immunostimulation nanoformula. The preparation method comprises the steps of: (1) providing a nanogold aqueous solution; (2) modifying the 3' terminal of CpG ODN (oligodeoxynucleotides) by using polyadenylic acid to obtain CpG ODN-polyA; and (3) assembling the CpG ODN-polyA to nanogold to form the multivalent immunostimulation nanoformula. The invention further provides the multivalent immunostimulation nanoformula obtained by the preparation method and application of the multivalent immunostimulation nanoformula. Compared with the traditional CpG ODN, the multivalent immunostimulation nanoformula has very high stability, no influence on cell activity, high cellular uptake rate, low administration dosage, good immunostimulation activity and remarkable effect during administration of low dosage.
Owner:SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
Eriocheir sinensis Crustin-1 gene and in-vitro recombination expression
InactiveCN101525617AAntibacterial agentsPeptide/protein ingredientsAureobasidium sp.Open reading frame
The invention relates to the technical field of molecular biology, in particular to eriocheir sinensis Crustin-1 gene clone and a recombination expression technology thereof. Crustin-1 gene of which the cDNA span is 821bp is cloned from eriocheir sinensis by using an expressed sequence tag (EST) technology and a 3' and 5' end rapid augmentation technology (RACE) and comprises a 315bp open reading frame, polyadenylic acid tailing signals and polyadenylic acid tails, 104 amino acids are coded, the length of a 5' non-coding region is 239bp, the length of a 3' non-coding region is 267bp, and the Crustin-1gene play an important role in the immune defense aspect of the eriocheir sinensis. The invention obtains eriocheir sinensis Crustin-1 protein by using the in-vitro recombination expression technology, the recombination protein has stronger bacteriostatic activity for gram-positive bacteria, the minimal inhibitory concentrations of the recombination protein for micrococcus luteus, bacillus subtilis, micrococcus tetragenus and bacillus thuringiensis are 0.12 mu m, 0.23 mu m, 0.46 mu m and 0.12 mu m respectively, and the recombination protein does not have obvious inhibiting function. The invention can lay the foundation for the disease control of the eriocheir sinensis, the gene assistant breeding and the development of feed additives.
Owner:INST OF OCEANOLOGY - CHINESE ACAD OF SCI
Mango ethylene receptor gene
ActiveCN106701785AReduce releaseDelayed release peakFruit and vegetables preservationPlant peptidesPolyadenylic acidRipening
The invention relates to the field of biotechnology, in particular to a mango ethylene receptor gene, which is named as MiETR1. Base sequences of the gene MiETR1 are as shown in SEQ ID NO:1 in a sequence table; the gene MiETR1 totally has 2570 base sequences including 113 3' untranslated regions, one polyadenylic acid tail and one opening decode regions with 2220 base sequences; an amino acid sequence of the gene MiETR1 is as shown in SEQ ID NO:2 in the sequence table. According to the mango ethylene receptor gene provided by the invention, the mango ethylene receptor gene MiETR1 is obtained through separating and cloning from mango, and an ethylene inhibitor is acted on the ethylene receptor gene MiETR1, so that an expression of the ethylene receptor gene MiETR1 can be remarkably inhibited, the ethylene release rate is reduced, the ethylene release peak is delayed, and the aim of delaying the ripening and senescence of the mango is achieved.
Owner:AGRI PROD PROCESSING INST GUANGXI ACADEMY OF AGRI SCI
MRNA and novel coronavirus mRNA vaccine containing same
PendingCN114480442AStable structureImprove expression efficiencySsRNA viruses positive-senseVirus peptidesCoronavirus vaccinationLiposome
The invention discloses a separated mRNA (messenger Ribonucleic Acid), which comprises an mRNA for coding a new coronavirus S1 protein, and the amino acid sequence of the S1 protein is as shown in SEQ ID NO: 2; the invention relates to a polyadenylic acid derivative which is characterized by further comprising one or more of the following (a)-(d): (a) a 5 '-cap structure, (b) 3'-polyadenylic acid, (c) 5 '-UTR and (d) 3'-UTR, also disclosed are DNA, compositions comprising the same, liposome nanoparticles, mRNA vaccines against new coronavirus, pharmaceutical compositions, and kits. The mRNA can be highly expressed in cells; the method is economical, efficient, safer and efficient, the structure is more stable, the protein expression efficiency is higher, and the new coronavirus S1 protein can be continuously expressed. When the liposome nano-particles / vaccines are prepared from the liposome nano-particles / vaccines, an enough protection effect can be achieved by using an extremely small dosage, the immunogenicity is reduced, and meanwhile, an organism immune system can be activated to generate humoral immunity and cellular immunity.
Owner:SHENZHEN GINO BIOTECHNOLOGY CO LTD
Enhancer for free circulating tumor DNA extraction, kit and method for extracting free circulating tumor DNA in peripheral blood
ActiveCN109371018AImprove extraction efficiencyIncrease productionDNA preparationYeastPolyadenylic acid
The invention discloses an enhancer for free circulating tumor DNA extraction, a kit and a method for extracting free circulating tumor DNA in peripheral blood, wherein the enhancer comprises 1-5 [mu]g / [mu]L of carrier RNA, 1-5 [mu]g / [mu]L of polyadenylic acid, 1-5 [mu]g / [mu]L of glycogen, 0-5 [mu]g / [mu]L of linear acrylamide, 1-5 [mu]g / [mu]L of yeast tRNA, 2-10 [mu]g / [mu]L of salmon sperm DNA and / or menhaden sperm DNA and water. The enhancer for the free circulating tumor DNA (ctDNA) extraction provided by the invention can be used in combination with a conventional lysed binding solution, awashing buffer, an eluant and the like, can be specifically bound to ctDNA and deposited, and is capable of significantly improving the extraction efficiency and the extraction yield of ctDNA.
Owner:BEIJING USCI MEDICAL DEVICES CO LTD
Expression vector for fusion expression of green fluorescent protein, construction method and use thereof
InactiveCN101463362AImprove the level ofEfficient exclusionFermentationVector-based foreign material introductionFusion Protein ExpressionOrigin of replication
The invention discloses an expression vector expressing green fluorescent protein by fusion and a construction method and application thereof. The vector is in ring shape , comprising a procaryon replication origin, two eucaryon replication origins and selective marker genes, and two green fluorescent protein gene expression cassettes; the green fluorescent protein gene expression cassettes, from upstream to downstream, consist of two promoters with the same transcription direction, junction fragments, green fluorescent protein encoding genes and polyadenylic acid AATAAA; the green fluorescent protein encoding gene lacks an initiation codon. The junction fragment comprises an EcoRV recognition sequence; the first place and the second place of the green fluorescent protein encoding gene from the end of 5' overlaps the last two places from the end of 5' of the EcoRV recognition sequence; the junction fragment comprises or does not comprise the initiation codon; when the junction fragment comprises the initiation codon, the initiation codon has a space of 2+3n or 1+3n from the first place from the end of 5' of the EcoRV recognition sequence, wherein, n is a positive integer. The vector comprises a green fluorescent protein gene, and can only express the green fluorescent protein on the condition that the gene is correctly expressed, therefore, the interference due to the expression of fluorescence by an initial vector can be effectively excluded.
Owner:CHINA AGRI UNIV
Avian origin promoter expression vector, construction method and use thereof
InactiveCN101481703AQuick buildIdeal connection efficiencyFermentationVector-based foreign material introductionOrigin of replicationNucleotide
The invention discloses an expression vector for an avian promoter and a construction method and an application thereof. The expression vector of the avian promoter is a ring-shaped vector, comprising a procaryon replication origin, a eucaryon replication origin, screening marker genes and a foreign gene expression box; wherein, the foreign gene expression box, from the upstream to the downstream, contains an LTR promoter, a junction fragment and polyadenylic acid tailing signals; the nucleotide sequence of the LTR promoter is the 1st to the 607th position counted from the end of 5' in a sequence 1 in a sequence table; the junction fragment contains EcoRV recognition sequence. With the pAPP-Vector of the invention adopted, the process to construct and express the recombinant vectors of the foreign genes is simple and convenient, rapid, economical and highly efficient; in addition, construction of the expression vectors of a large number of the foreign genes can be accomplished very rapidly; moreover, the pAPP-Vector of the invention can play an important role in studies on transgenic animals.
Owner:CHINA AGRI UNIV
Eukaryotic cell system for detecting integrase phi-C31 functions and preparation method thereof
InactiveCN1827781AImprove accuracyGood repeatabilityMicrobiological testing/measurementBiotechnologyΦc31 integrase
The invention relates the eukaryotic cell system and method for checking integrated enzymeªÁC31 function, belonging to biotechnology and causal treatment technology field. The method comprises the following steps: after recombination report PB [EGFP] transfer dyeing the embryon kidney cell system, screening checking system, choosing monoclonal cell, getting the product, the PB[EGFP] using the pEGFP-N2 as carrier, after enhancing CMVIE, sequence inserting specificity sequence attP and attB, nd then adding growth factor polyadenylic acid. The cell system can estimate the integrated enzymeªÁC31 expressed by transfer dyeing, transduction and conversion approach, and the activity and function of its mutant. The checking system has the advantages of simple operation, high accuracy, and good repeatability. The system is the important quantitative estimation system that integrated enzymeªÁC31 is in the application of mammalian cytology.
Owner:FUDAN UNIV
Method for preparing mRNA and application of the mRNA in tumor treatment
PendingCN110331147AImprove translation efficiencyGrowth inhibitionPeptide/protein ingredientsNGF/TNF-superfamilyPolyadenylic acidA-DNA
The invention discloses a method for preparing an mRNA and application of the mRNA in tumor treatment. The invention further discloses a nucleic acid molecule, which comprises a promoter, a transcribable segment A, a transcribable segment B, a transcribable segment C and a polyadenylic acid sequence in turn. Driven by the promoter, the transcribable segment A, the transcribable segment B, the transcribable segment C and the polyadenylic acid sequence can be co-transcribed to produce a common transcript; the segment A encodes a 5'-end untranslated region in the common transcript; the segment Cencodes a 3'-end untranslated region in the common transcript; the transcribable segment A is a DNA molecule shown as sequences 1-4; and the transcribable segment C is a DNA molecule shown as sequences 5-8. The mRNA molecule is obtained by in vitro transcription adopting the nucleic acid molecule as a template. The nucleic acid molecule or mRNA molecule provided can be used for preparing medicinesfor treating various tumors.
Owner:SUZHOU GENEPHARMA +1
High-throughput identification method of internal ribosome entry site (IRES) elements in various source cell samples
ActiveCN110819707AImprove accuracyAvoid Consistency EffectsMicrobiological testing/measurementNucleic acid vectorCDNA libraryBinding site
The invention discloses a high-throughput identification method of internal ribosome entry site (IRES) elements in various source cell samples. The method comprises the following steps: a full-lengthcDNA library is obtained from a sample by utilizing oligonucleotides designed for an mRNA chain 5'- terminal cap structure and a 3'-terminal polyadenylic acid tail through reverse transcription; the cDNA library is fragmented by utilizing TN5 transposase and a designed nucleic acid fragment containing a special linker sequence; active IRES elements are screened by utilizing a lentivirus reporter gene plasmid vector containing red fluorescent proteins, green fluorescent proteins and special elements, and false positive results caused in a translation process are eliminated; and the active IRESelements are identified by utilizing a long reading and long sequencing technology and false positive results caused by a promoter and variable shearing in the transcription process are eliminated. The method pushes high-throughput identification of IRES elements to clinical application, so that a role played by the IRES elements in a process of disease occurrence and development can be deeply studied.
Owner:XIANGYA HOSPITAL CENT SOUTH UNIV
Hemoglobin gene of macrobrachium nipponensis, cloning method of hemoglobin gene, and preparation method of recombinant protein
InactiveCN104073501AOxygen-carrying functionImprove antibacterial propertiesAntibacterial agentsPeptide/protein ingredientsOpen reading frameMacrobrachium nipponense
The invention discloses hemoglobin gene of macrobrachium nipponensis, a clone method of the hemoglobin gene, and a preparation method of recombinant protein. The hemoglobin gene of macrobrachium nipponensis is cloned through an expressed sequence tag technology and 3'and 5' rapid amplification cDNA end (RACE), wherein the overall length of the cDNA is 1201bp, the gene has a 579bp open reading frame and encodes 193 amino acids, 5' noncoding region is 199bp and 3' noncoding region is 420bp, and the gene has polyadenylic acid tailing signal and polyadenylic acid tail. The gene plays an important role in immune defense of macrobrachium nipponensis. The hemoglobin gene of macrobrachium nipponensis is obtained by in vitro recombination expression; the recombinant protein not only can carry oxygen, but also has bacteriostasis, thereby providing theoretical basis for disease control and gene-assisted breeding of macrobrachium nipponensis,and development of feed additives for macrobrachium nipponensis.
Owner:FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
Chlamys Farreri H2A gene clone and N terminal expression technology
The invention relates to a scallop protein H2A gene clone and the monoclonal expression technique of N end antibiotic polypeptide in the field of molecular biology technique. It uses isogenesis clone technique to obtain the scallop group protein H2A gene group total length; the gene DNA has a 375bp open reading frame with the coding 125 amino acids, 3' non-coding area comprises two ending signals which are stem ring structure and polyadenylic acid rear signal. It uses pPIC9K expressing carrier to express the protein N end 30aa in Pichia pastoris, the expressing product has bacterial inhibition to the Micrococcus luteus and the Vibrio splendidus.
Owner:INST OF OCEANOLOGY - CHINESE ACAD OF SCI
Detection method of single-stranded micro ribonucleic acid
InactiveCN101792806AEasy to detectQuick checkMicrobiological testing/measurementFluorescence/phosphorescenceSpecific detectionPolyadenylic acid
The invention relates to a detection method of single-stranded micro ribonucleic acid (miRNA), belonging to the technical field of biological detection. Cells are used or RNA (containing miRNA) is organized, wherein a polyadenylic acid tail is added to the 3' terminal of miRNA by poly (a) polymerase, then reverse transcription primers are used for reverse transcription to obtain single-stranded deoxyribonucleic acid (ssDNA) complementary to the RNA, finally the miRNA related sequences and partial complementary sequences of the reverse transcription primers are respectively used as the upperstream and downstream primers for miRNA detection and a specific detection system (containing SYBR Green I fluorescent dye) is used for detecting miRNA. The method has the advantages of simpleness, convenience, rapidness and suitability for detection of a large number of samples. The method can be used for expression analysis, cloning research and clinical sample test of miRNA.
Owner:TIANGEN BIOTECH BEIJING
Attenuated virus of flavivirus and application of attenuated virus
The present invention relates to an attenuated virus of a flavivirus and application of the attenuated virus. The attenuated virus comprises a polyadenylic acid (poly(A)) sequence, wherein the poly(A) is used for replacing a part of nucleotide sequence of a 3' untranslated region (3' UTR) of the yellow disease virus, so that the 3' untranslated region (3' UTR) of the attenuated yellow disease virus obtained after the part of nucleotide sequence of the yellow disease virus is replaced at least retains a 3'-end stem-loop region (3' SL). The attenuated virus has growth characteristics similar to those of a wild type virus, and is high in titer and good in hereditary stability. Moreover, the attenuation is fully attenuated, the safety is high, the generation of high-level neutralizing antibody titer can be induced by single immunization, the attack of high-dose wild-type viruses can be resisted, complete immune protection can be provided, and the attenuated vaccine strain can be used for preparing safe and effective attenuated vaccine strains.
Owner:湖北彰骏生物科技有限公司
RAAV vector for expressing antibody IgG1 and application of rAAV vector
PendingCN114107390AQuick buildSerum immunoglobulinsImmunoglobulins against cell receptors/antigens/surface-determinantsT cellPromoter
The invention discloses an rAAV vector for expressing an antibody IgG1 and application of the rAAV vector. The ITR core expression element of the rAAV vector is composed of a broad-spectrum strong promoter CMV, an IgG1 heavy chain secretion signal peptide sequence, an IgG1 heavy chain coding region, a connecting polypeptide coding sequence, an IgG1 light chain secretion signal peptide sequence, an IgG1 light chain coding region and a human serum albumin polyadenylic acid sequence. The heavy chain variable region (VH) and the light chain variable region (VL) can be conveniently and quickly operated through the restriction enzyme cutting sites on the rAAV vector. The antibody IgG1-rAAV obtained by packaging is used for infecting HEK293T cells or C57 mice and the like cultured in vitro, and various antibodies IgG1 with different antigen recognition activities can be expressed.
Owner:INNOVATION ACAD FOR PRECISION MEASUREMENT SCI & TECH CAS
Mosaic tumor specific gene expression regulation kit and application thereof
InactiveCN102051358AStrong specificityImprove targeted lethalityGenetic material ingredientsViruses/bacteriophagesPolyadenylic acidTumor therapy
The invention belongs to the technical field of biomedicines, and in particular provides a mosaic tumor specific gene expression regulation kit, which comprises a tumor specific gene promoter, a 5' untranslated region of a tumor specific translational control component, a polyclonal site, a gene expression enhancer and a polyadenylic acid signal region, wherein the tumor specific gene promoter, the 5' untranslated region of the tumor specific translational control component, the gene expression enhancer and the polyadenylic acid signal region are connected in turn; and the polyclonal site is positioned between the 5' untranslated region of the tumor specific translational control component and the polyadenylic acid signal region and is independent of the gene expression enhancer. The mosaic tumor specific gene expression regulation kit effectively regulates the expression of tumor therapy genes to limit the genes in tumor cells, and obviously improves the specificity of the expression of the therapy genes so as to effectively improve the safety of tumor gene therapy and reduce the harm of the tumor gene therapy to normal tissues and cells.
Owner:FUDAN UNIV
Wheat gene tafus3 and its application
InactiveCN105713909BHigh expressionResilience ImprovementPlant peptidesGenetic engineeringBiotechnologyTriticeae
The invention relates to the technical field of plant genetic engineering, in particular to a wheat stress resistance gene TaFUS3 and its application. The wheat stress-resistance-related gene TaFUS3 of the present invention is isolated from the ear-resistant germination variety "Huaimai 0360" and the easy-ear germination variety "Zhongyou 9507". The full-length cDNA of the gene is 1461 bp, including 47 bp of 5 ′-non-coding region; 484 bp 3′-non-coding region; 906 bp coding region, terminator, and 28 bp polyadenylation tail, the protein encoded by the gene is a full-length protein consisting of 301 amino acids Composed of B3 domain-containing proteins. The wheat stress-resistance-related gene TaFUS3 of the present invention can be applied to seed development and dormancy, stress-resistance response, panicle germination resistance, stress-resistant breeding, gene chip production, and the design and synthesis of new stress-resistance-related genes has high practical application value.
Owner:HENAN AGRICULTURAL UNIVERSITY
Chlamys Farreri H2A gene clone and N terminal expression technology
InactiveCN100500845CAntibacterial agentsPeptide/protein ingredientsPichia pastorisOpen reading frame
The invention relates to a scallop protein H2A gene clone and the monoclonal expression technique of N end antibiotic polypeptide in the field of molecular biology technique. It uses isogenesis clone technique to obtain the scallop group protein H2A gene group total length; the gene DNA has a 375bp open reading frame with the coding 125 amino acids, 3' non-coding area comprises two ending signals which are stem ring structure and polyadenylic acid rear signal. It uses pPIC9K expressing carrier to express the protein N end 30aa in Pichia pastoris, the expressing product has bacterial inhibition to the Micrococcus luteus and the Vibrio splendidus.
Owner:INST OF OCEANOLOGY - CHINESE ACAD OF SCI
Silkworm PABP (polyadenylic acid bonding protein) bonding protein interacting factor gene BmPaip1 as well as recombinant expression vector and application thereof
ActiveCN103060334BImprove expression levelImprove translation efficiencyFermentationVector-based foreign material introductionOpen reading frameExon
The invention discloses a silkworm PABP (polyadenylic acid bonding protein) bonding protein interacting factor gene BmPaip1. The nucleotide sequence of the gene BmPaip1 is shown as SEQIDNO.3, the open reading frame of the gene BmPaip1 is 1194bp long, composed of eight exons and codes 398 amino acids, theoretical protein molecular weight is 44kDa, and isoelectric point pI is equal to 5.02; and SMART online software analysis shows that the bits from 100 to 398 in the amino acid sequence of the gene BmPaip1 of a silkworm are highly homologous with an eIF4G protein of an eucaryon, and expression detection in insect cells shows that expression level of a reporter gene LUC (luciferase) can be obviously improved in cells genetically modified by the gene BmPaip1 of the silkworm, so that the gene BmPaip1 of the silkworm can be combined with eIF4E and eIF4A to form a translation initiation complex and can be used for improving translation efficiency of an exogenous gene.
Owner:SOUTHWEST UNIV
Swine promoter protein expression vector and construction method and application thereof
InactiveCN101948872BImprove the level ofIdeal connection efficiencyVector-based foreign material introductionOrigin of replicationPolyadenylic acid
The invention discloses a swine promoter protein expression vector and a construction method and application thereof. The expression vector is annular and comprises a pronucleus replication origin, an eukaryon replication origin, a selection marker gene and an exogenous gene expression cassette, wherein the exogenous gene expression cassette consists of a transcription promoter, a junction fragment and a polyadenylic acid tailing signal sequentially from upstream to downstream; and the transcription promoter is an MLP6 promoter. The expression vector can efficiently express exogenous proteins in a swine cell, contains the eukaryon replication origin and can be proliferated greatly in a cell of an expression T antigen, so that protein expression level is raised. A recombinant vector of an expression exogenous gene is constructed by homologous recombination, so that the conventional steps such as enzyme cutting, linking and the like are avoided and 2 to 3 steps are saved compared with the conventional linking process. A vector recombination process for constructing the expression exogenous gene by using the vector of the invention is simple, convenient, rapid, economical and efficient and the construction of the expression vectors of a large number of exogenous genes can be finished rapidly.
Owner:HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
Swine promoter protein expression vector and construction method and application thereof
InactiveCN101948872AImprove the level ofIdeal connection efficiencyVector-based foreign material introductionOrigin of replicationPolyadenylic acid
Owner:HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
Prawn virus expression system for delivering and expressing exogenous genes in prawns
The invention provides a prawn virus expression system for delivering and expressing an exogenous gene in a prawn cell, which comprises a recombinant plasmid for expressing the exogenous gene in the prawn cell, the recombinant plasmid comprises genome DNA (deoxyribonucleic acid) of prawn IHHNV virus, a promoter PH (potential of hydrogen) of insect polyhedrosis protein gene, a polyclone restriction enzyme cutting site MCS (multi-cloning restriction enzyme cutting site) for inserting an exogenous gene and an SV40 polyadenylic acid signal pA. The expression system further comprises an insect baculovirus recombinant expression plasmid and an insect Sf9 packaging cell, wherein the gene of the WSSV envelope protein VP28 is inserted into the insect baculovirus recombinant expression plasmid. The prawn virus expression system provided by the invention can efficiently express exogenous genes in prawn cells, and all plasmid DNA genetic information in the expression system is free out of chromosomes of the prawn cells and cannot be integrated into genomes of the prawn cells, so that the biological safety of the system is improved.
Owner:OCEAN UNIV OF CHINA
Eukaryon expression shuttle vector, construction method and application
InactiveCN101343639BQuick buildEfficient constructionVector-based foreign material introductionOrigin of replicationShuttle vector
The invention discloses an eucaryon expressing shuttle vector as well as a constructing method and applications thereof. The eucaryon expressing shuttle vector adopts an annular vector which comprises a prokaryon replication origin, two eucaryon replication origins, selective marker genes and a foreign gene expressing box; the foreign gene expressing box is composed of two promoters sharing an identical duplicating direction, connecting segments and polyadenylic acid tailing signals from upstream to downstream sequentially; and the connecting segment contains an EcoRV recognition sequence. The process of constructing the recombinant vector of expression foreign gene through the eucaryon expressing shuttle vector is convenient, quick, economical and highly efficient, and the vector contains two eucaryon replication origins, therefore, the vector can be greatly appreciated in cells expressing T antigen, and the protein expressing level can be improved. The eucaryon expressing shuttle vector has broad application prospect.
Owner:CHINA AGRI UNIV
Cell-free translation system, method and product
PendingCN113409887AOvercome inability to expressOvercome the problem of long expression cycle and low efficiencyProteomicsGenomicsProtein targetCell free
The invention relates to the field of molecular biology, in particular to a cell-free translation system, method and product. The translation system comprises mRNA of 5'cap and 3 'polyadenylic acid, an amino acid mixture, an energy molecule and a mixture of mRNA translation enzyme sources; the energy molecule comprises phosphocreatine, ATP (adenosine triphosphate) and guanosine triphosphate (GTP). Through mRNA transcription, in-vitro transcription and gel electrophoresis analysis, the translation system can rapidly obtain a large amount of target proteins and has the following characteristics: an extracellular translation technology is adopted to overcome the problem that a prokaryotic cell expression part gene cannot be expressed; the problems of long expression cycle and low efficiency of eukaryotic cells are solved by adopting an extracellular translation technology; the problems of low activity and poor efficiency of the original extracellular translation technology are solved by adopting a strategy of adding energy micromolecules such as phosphocreatine; and the stability, the receptor binding speed and the overall translation speed are enhanced by adopting mRNA of 5 ' cap and 3' polyadenylic acid.
Owner:INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
A mango ethylene receptor gene
ActiveCN106701785BReduce releaseDelayed release peakFruit and vegetables preservationPlant peptidesPolyadenylic acidRipening
The invention relates to the field of biotechnology, in particular to a mango ethylene receptor gene, which is named as MiETR1. Base sequences of the gene MiETR1 are as shown in SEQ ID NO:1 in a sequence table; the gene MiETR1 totally has 2570 base sequences including 113 3' untranslated regions, one polyadenylic acid tail and one opening decode regions with 2220 base sequences; an amino acid sequence of the gene MiETR1 is as shown in SEQ ID NO:2 in the sequence table. According to the mango ethylene receptor gene provided by the invention, the mango ethylene receptor gene MiETR1 is obtained through separating and cloning from mango, and an ethylene inhibitor is acted on the ethylene receptor gene MiETR1, so that an expression of the ethylene receptor gene MiETR1 can be remarkably inhibited, the ethylene release rate is reduced, the ethylene release peak is delayed, and the aim of delaying the ripening and senescence of the mango is achieved.
Owner:AGRI PROD PROCESSING INST GUANGXI ACADEMY OF AGRI SCI
Application of attenuated yellow disease virus in oncolysis
PendingCN113717953ACause toxic side effectsSmall side effectsVirusesHydrolasesDiseaseGenus Dependovirus
The present invention relates to an oncolytic virus and use thereof, the oncolytic virus comprising an attenuated flavivirus, the attenuated flavivirus comprising a polyadenylic acid (poly (A)) sequence, wherein the poly (A) sequence is used for replacing a part of nucleotide sequence of a 3' untranslated region (3' UTR) of the yellow disease virus, so that the 3' untranslated region (3' UTR) of the attenuated yellow disease virus obtained after the part of nucleotide sequence of the yellow disease virus is replaced at least retains a 3'-end stem-loop region (3' SL). In-vitro cell experiments prove that the oncolytic virus disclosed by the invention can infect and kill various tumor cells, but has small influence on normal cells; and a xenotransplantation tumor model proves that the oncolytic virus disclosed by the invention can inhibit the growth of various tumor cells in vivo, and a safe and efficient anti-cancer therapeutic agent with small toxic and side effects can be provided for clinical tumor treatment.
Owner:BEIJING SHUNLEI BIOTECHNOLOGY CO LTD +1
Macrobrachium japonicus hemoglobin gene and its cloning method and recombinant protein preparation method
InactiveCN104073501BOxygen-carrying functionImprove antibacterial propertiesAntibacterial agentsPeptide/protein ingredientsOpen reading frameFeed additive
The invention discloses hemoglobin gene of macrobrachium nipponensis, a clone method of the hemoglobin gene, and a preparation method of recombinant protein. The hemoglobin gene of macrobrachium nipponensis is cloned through an expressed sequence tag technology and 3'and 5' rapid amplification cDNA end (RACE), wherein the overall length of the cDNA is 1201bp, the gene has a 579bp open reading frame and encodes 193 amino acids, 5' noncoding region is 199bp and 3' noncoding region is 420bp, and the gene has polyadenylic acid tailing signal and polyadenylic acid tail. The gene plays an important role in immune defense of macrobrachium nipponensis. The hemoglobin gene of macrobrachium nipponensis is obtained by in vitro recombination expression; the recombinant protein not only can carry oxygen, but also has bacteriostasis, thereby providing theoretical basis for disease control and gene-assisted breeding of macrobrachium nipponensis,and development of feed additives for macrobrachium nipponensis.
Owner:FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
Wheat disease-resistant gene Lr-L1 and its use
InactiveCN1280415CEasy to controlDisease resistantPlant peptidesFermentationPoly-A RNAAgricultural science
The invention relates to the technical field of plant genetic engineering, in particular to a wheat disease resistance gene Lr-L1 and its application. The full-length cDNA of the wheat disease-resistant gene Lr-L1 of the present invention is 2326bp, including a 5'-noncoding region of 135bp, a 3'-noncoding region of 249bp, a coding region of 1911bp, a terminator, and a polyadenylic acid of 28bp Tail, the 3′-noncoding region contains three typical tailing signal sequences, which can strengthen the tailing process after gene transcription. The wheat disease-resistant gene Lr-L1 of the present invention can be inserted into the downstream of different types of promoters to construct a plant expression vector, and the plant disease resistance can be improved through genetic engineering technology, and the gene can also be cloned for the production of wheat gene chips. In the research of wheat disease resistance and so on.
Owner:HENAN AGRICULTURAL UNIVERSITY
Expression vector for fusion expression of green fluorescent protein, construction method and use thereof
InactiveCN101463362BImprove the level ofEfficient exclusionFermentationVector-based foreign material introductionOrigin of replicationStart codon
The invention discloses an expression vector expressing green fluorescent protein by fusion and a construction method and application thereof. The vector is in ring shape , comprising a procaryon replication origin, two eucaryon replication origins and selective marker genes, and two green fluorescent protein gene expression cassettes; the green fluorescent protein gene expression cassettes, fromupstream to downstream, consist of two promoters with the same transcription direction, junction fragments, green fluorescent protein encoding genes and polyadenylic acid AATAAA; the green fluorescent protein encoding gene lacks an initiation codon. The junction fragment comprises an EcoRV recognition sequence; the first place and the second place of the green fluorescent protein encoding gene from the end of 5' overlaps the last two places from the end of 5' of the EcoRV recognition sequence; the junction fragment comprises or does not comprise the initiation codon; when the junction fragment comprises the initiation codon, the initiation codon has a space of 2+3n or 1+3n from the first place from the end of 5' of the EcoRV recognition sequence, wherein, n is a positive integer. The vector comprises a green fluorescent protein gene, and can only express the green fluorescent protein on the condition that the gene is correctly expressed, therefore, the interference due to the expression of fluorescence by an initial vector can be effectively excluded.
Owner:CHINA AGRI UNIV
Chromosome landing pads and related uses
The present invention provides methods for stably integrating and / or expressing one or more recombinant polynucleotides in a host cell. The recombinant polynucleotide is usually integrated into the host genome at some natural chromosomal integration site. The integration can be mediated by homologous recombination or with hybrid recombinases targeted to specific chromosomal locations. Stable integration and strong transcriptional activity of foreign genes are supported by native chromosomal integration sites in the host cell, which are located within or near specific genes in the CHO genome, the ankyrin 2 gene (Ank2), cleavage and polyadenylation-specific factor 4 gene (Cpsf4), C-Mos gene and Nephrocystin-1 / Mal gene. Also provided are methods and nucleic acid molecules for the insertion of site-specific recombination sequences (chromosomal landing pads) into these specific chromosomal locations. Further provided are engineered host cells containing chromosomal landing pads or transgenes integrated into natural chromosomal integration sites. Further provided are methods and compositions (eg, kits) for integrating and expressing heterologous polynucleotides in host cells with inserted chromosomal landing pads.
Owner:THE SCRIPPS RES INST
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