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44 results about "LacZ Genes" patented technology
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Genetic engineering bacterium for producing monophosphoryl lipid A as well as construction method and application thereof
InactiveCN102399736AReduce manufacturing costConducive to large-scale industrial productionBacteriaMicroorganism based processesMicrobiologyBacterial strain
The invention discloses a genetic engineering bacterium for producing a monophosphoryl lipid A as well as a construction method and application thereof, belonging to the field of a genetic engineering. The genetic engineering bacterium is E.coli W3110 delta lacI lacZ::FnlpxE, wherein the lacI takes place a deletion mutation to lose activity and an expression FnlpxE gene is knocked into the lacZ gene. According to the bacterial strain constructed in the invention, on the basis of keeping a lipid structure single, the production cost is also reduced; no exogenous resistance genes are introduced into the bacterial strain so that the industrial production of the bacterial strain is easier to realize in a large scale.
Owner:JIANGNAN UNIV
Genetically engineered bacterium of colon bacillus for producing arabinoside-cytidine monophosphate lipoid A, and application thereof
The invention discloses a genetically engineered bacterium of a colon bacillus for producing arabinoside-cytidine monophosphate lipoid A. The genetically engineered bacterium is characterized in that a deletion mutation deactivation is carried out on lacz genes in a genome of the genetically engineered bacterium; and express exogenous pagL and lpxE and pagP genes are implanted into the lacz genes. According to the invention, the formed bacterial strain cannot produce a pathogenic bacterium and is free of the use of an inductive agent during a production process. Therefore, the genetically engineered bacterium is suitable for mass production of MPL adjuvants in earlier stages.
Owner:JIANGNAN UNIV
Recombinant escherichia coli for synthesizing lactoyl N-neotetraose and construction method and application of recombinant escherichia coli
The invention discloses recombinant escherichia coli for synthesizing lactoyl N-neotetraose and a construction method and application of the recombinant escherichia coli. The recombinant escherichia coli are obtained by knocking out a beta-galactosidase lacZ gene, a glucosamine-6-phosphodeaminase nagB gene, a UDP-acetylglucosamine epitope isomerase wecB gene and a UDP-glucose dehydrogenase ugd gene in the host genome of escherichia coli and overexpressing a lactose transportase lacY gene, beta-1, 3-N-glucosaminidase lgtA gene and a beta-1, 4 galactosyltransferase lgtB gene on the genome. The yield of lactoyl-N-neotetraose synthesized by recombinant escherichia coli reaches 985 mg / L, and a foundation is laid for further metabolic engineering transformation of escherichia coli to produce lactoyl-N-neotetraose.
Owner:JIANGNAN UNIV
A recombinant fowlpox virus transfer vector expressing a fowl adenovirus serotype-4 fiber2 gene, a constructing method thereof and applications of the transfer vector
ActiveCN107475296AEliminate conflictImprove efficiencyViral antigen ingredientsVirus peptidesVector vaccineEmbryo
The invention belongs to the technical field of biology, and mainly relates to a recombinant fowlpox virus transfer vector expressing a fowl adenovirus serotype-4 (FADV4) fiber2 gene, a constructing method thereof and applications of the transfer vector. A pMD19T-Simple vector is adopted as a base of the transfer vector. The FADV4 fiber2 gene, a lacz gene, and fowlpox virus genome replicated non-essential fragments which are LTYB and RTYB used for homologous recombination are inserted in a TA cloning site. The method includes constructing a plasmid pMD-TYB; constructing a plasmid pMD22; constructing a plasmid pMD22-lacz; constructing an intermediate vector pMD22-TYB-lacz; amplifying the FADV4 fiber2 gene; constructing a recombinant fowlpox virus transfer vector pMD22-TYB-lacz-F4; subjecting a chicken embryo Fibroblast to cotransfection with the transfer vector pMD22-TYB-lacz-F4 and a fowlpox virus; performing identification to select a positive product; performing subculture continuously; and identifying expression effects of the recombinant fowlpox virus. The recombinant fowlpox virus transfer vector constructed by the method lays a foundation for development of an efficient recombinant fowlpox virus genetic engineering living-vector vaccine expressing the fowl adenovirus serotype-4.
Owner:WENS FOODSTUFF GRP CO LTD
PC series plasmid as well as construction method and application thereof
ActiveCN102352371BLarge capacitySmooth extractionVector-based foreign material introductionAntibiotic YGene engineering
Owner:GENEWIZ INC SZ
Rapid screening system for Lactococcus lactis with knocked-in exogenous genes as well as construction method and application of rapid screening system
InactiveCN105349565AReduce workloadShorten the timeBacteriaMicrobiological testing/measurementStaphylococcus lactisBiology
The invention provides a rapid screening system for Lactococcus lactis with knocked-in exogenous genes. The screening system comprises temperature-sensitive plasmids and the Lactococcus lactis, wherein the temperature-sensitive plasmids contain his gene segments and temperature-sensitive replicon-erythromycin resistance gene (Ts-Emr) segments; chromosomes of the Lactococcus lactis contain lacZ gene expression segments. lacZ genes of the Lactococcus lactis cannot be expressed when the exogenous genes are knocked in successfully, so that white colonies are shown on a solid medium containing X-Gal, otherwise, blue colonies are shown, the change from the blue colonies to the white colonies can be directly observed on the solid medium, the screening workload can be greatly reduced, and screening time can be greatly shortened.
Owner:ARMY MEDICAL UNIV
PC series plasmid as well as construction method and application thereof
ActiveCN102352371ALarge capacitySmooth extractionMicrobiological testing/measurementVector-based foreign material introductionAntibiotic YGene engineering
The invention discloses a pC series plasmid as well as a construction method and application thereof, wherein the pC series plasmid comprises a rep gene, a rop gene, a lacZ gene and an antibiotics resistance gene which are sequentially connected with one another end to end. The pC series plasmid has the characteristics of small molecular weight, large capacity, low copy number, high stability andthe like, can be used for instable genes with complicated structures and repeated sequence, cloning, screening and sequencing long section of genes, genome construction and other gene engineering fields.
Owner:GENEWIZ INC SZ
Recombinant escherichia coli for synthesizing 2 '-fucosyllactose by using mannose and application of recombinant escherichia coli
PendingCN114276971AImprove conversion rateImprove carbon source utilizationBacteriaMicroorganism based processesEscherichia coliHigh mannose
The invention relates to a method for constructing recombinant escherichia coli, synthesizing GDP-fucose by using mannose, further producing 2 '-fucosyllactose and improving the utilization rate of mannose, and belongs to the field of microbial metabolism engineering. The engineering bacterium takes escherichia coli as a starting strain, a Plac promoter sequence and lacI and lacZ genes in a lac operon sequence are knocked out, wcaG, gmd and lacy genes are overexpressed on 1acI and lacZ gene loci, then a phosphomannose isomerase coding gene manA is knocked out on a genome, a phosphomannose mutase coding gene manB and alpha-(1, 2, 3, 4, 5, 6, 7, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8, 8 and (2) obtaining a fucosyl transferase coding gene futC and a mannose-1-phosphate guanine transferase coding gene manC. According to the fermentation strategy for producing the 2 '-fucosyllactose through the de novo synthesis route of the 2'-fucosyllactose, the utilization rate of a carbon source for producing the 2 '-fucosyllactose through escherichia coli fermentation can be greatly increased.
Owner:TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY +1
Orf virus virulence gene VIR-deleted (interferon-resistance gene deleted) mutant and its preparing method and application
InactiveCN104878043ALow toxicityImproving immunogenicityAntiviralsViruses/bacteriophagesEnzyme digestionShuttle vector
The invention relates to an orf virus virulence gene VIR-deleted (interferon-resistance gene deleted) mutant and its preparing method and application. The preparing method includes: cloning a gene segment containing an ORFV-SHZ1 VIR virulence gene and a virulence gene by means of PCR (polymerase chain reaction), deleting the VIR virulence gene by means of enzyme digestion, inserting an expression cassette carrying vaccinia virus late promoter P11 and a LacZ reporter gene into a deletion site, and building a recombinant shuttle vector using the LacZ gene as a selection marker; transfecting the recombinant shuttle vector to ORFV infected Vero cells to allow homologous recombination, and acquiring the VIR virulence gene deleted recombinant virus ORFV-DeltaVIR-LacZ by means of plaque screening and purifying. The mutant has the advantages that the ORFV virulence is lowered quickly by means of gene engineering, technical support is provided for the research and development of ORF vaccines, the preparing method features short production cycle, avoidance of domestication, operational convenience and quickness and the like, and the preparing method is widely applicable.
Owner:SHIHEZI UNIVERSITY
Fowl pos virus expression vector P12 18 and its construction process
The present invention relates to fowl pos virus expression vector constructing technology, and is especially fowl pos virus expression vector p12-18 and its construction process. The fowl pos virus expression vector p12-18 constructing process includes the following steps: screening newly copied nonessential regions of fowl pos virus and amplifying fowl pos virus to copy two flanking regions of nonessential region FPV12-18; cloning these two flanking regions and marker gene expressing frame P11-LacZ successively into vector pBluescript SK(-), cotransfecting chick embryo fibroblast with the obtained vector pPl2 and FPV of 282E4 strain, and screening the recombinant virus with the marker gene of colibacillus beta-galactosidase LacZ gene to obtain recombinant virus vector; and finally inserting the exogenous gene promoter Ps into the vector to obtain fowl pos virus expression vector. The present invention can obtain recombinant fowl pos virus vaccine with high capacity.
Owner:YANGZHOU UNIV
Construction method and application of bacterial promoter reporter vector
InactiveCN110628799AReliable resultsGood repeatabilityMicrobiological testing/measurementBiological material analysisPromoter activityDNA fragmentation
The invention discloses a construction method and application of a bacterial promoter reporter vector. The method includes the following steps: first constructing a vector containing beta-galactosidase lacZ gene, inserting a target gene promoter into the upstream of the lacZ gene in the vector so as to obtain a reporter vector for detecting the activity of the gene promoter, introducing the reporter vector into to-be-tested strains, and inferring the change of the activity of the target gene promoter by detecting and comparing the activity of the strains and the beta-galactosidase. The reporter vector of the present invention can be used to detect the activity of gene promoters in bacteria and determine the strength of gene expression, and can also be used to detect the presence or absenceof promoter activity of unknown DNA fragments in bacteria, and to determine the location of the gene promoters with low operation cost and stable and reliable result.
Owner:武汉博欧特生物科技有限公司
Fowl pox virus double-gene expression carrier (PG7.5N)
InactiveCN101220374AFlexibleImprove expression efficiencyOrganic active ingredientsGenetic material ingredientsOrigin of replicationAgricultural science
The invention provides a fowlpox virus double-gene expression vector, which has a nucleotide sequence that is shown by SEQ ID NO.1. The invention includes a promoter P7.5, an LacZ gene, multiple cloning sites which are located on the LacZ gene, a rare restriction enzyme cutting site Not1, homologous arms F11L1 and F11L2 of the fowlpox virus, an Amp <R> resistance marker gene and an origin of replication (ori). The double-gene expression vector constructed by the invention can reduce the length of the vector to the maximum extent, thus a larger exogenous gene can be contained; the number of available restriction enzyme cutting sites is increased so as to facilitate the cloning of different exogenous genes; the promoter at an insertion site Not1 is not limited and the promoter which is matched with the exogenous gene can be freely chosen so as to improve expression efficiency; furthermore, the invention adopts a reverse expression method so as to improve the expression efficiency of the exogenous gene.
Owner:SOUTH CHINA AGRI UNIV
Prokaryotic promoter reporting system based on lacZ gene and pUC replicon, and construction method and application thereof
ActiveCN111235172AEfficient identificationImprove stabilityBacteriaMicroorganism based processesBiotechnologyReplicon
The invention discloses a prokaryotic promoter reporting system based on a lacZ gene and a pUC replicon. The reporting system comprises the pUC replicon derived from pFLX107, a Cat gene, an rrnbT1T2,a lambda t0 transcription terminator, and the lacZ gene derived from a plasmid pRCL, wherein ClaI, SacI and NdeI sites in the lacZ gene are subjected to silent mutation. The invention further discloses a construction method and application of the prokaryotic promoter reporting system. The size of a plasmid pFGH06 constructed by the invention is 4949 bp; the background activity of the plasmid pFGH06 under a culture condition of 28 DEG C is only 12.2 + / - 0.6, which is remarkably lower than the background activity (21.3 + / - 1.7) of the low-copy reference plasmid pRCL; the plasmid pFGH06 is applied to cloning and activity determination of an inducible promoter araBAD and a constitutive promoter rpsM; and when the plasmid pFGH06 is applied to promoter screening in a simulated mode, 100% recognition of a target promoter can be achieved through blue-white selection.
Owner:YANGZHOU UNIV
New lactic acid bacteria
The invention relates to a polynucleotide comprising a lacZ gene (lacZFS) encoding a [beta]-galactosidase characterized by a particular profile regarding its efficiency of hydrolysis of lactose. The invention is also directed to a Streptococcus thermophilus strain comprising a lacZFS allele and bacterial composition thereof, and their use to obtain fermented milk not undergoing post-acidification.
Owner:DUPONT NUTRITION BIOSCIENCES APS
Sheep aphthous virus virulence gene vir deletion mutant strain and preparation method and application thereof
InactiveCN104878043BLow toxicityImproving immunogenicityAntiviralsViruses/bacteriophagesVirulent characteristicsShuttle vector
The invention relates to a mutant strain of the virulence gene VIR of aphthous ulcer virus and its preparation method and application. The preparation method comprises the method of cloning the gene fragment containing the VIR virulence gene of ORFV-SHZ1 strain and the flanking sequence by using the PCR method, and deleting it by the enzymatic cutting method. Remove the VIR virulence gene, insert the expression cassette with the late promoter P11 of vaccinia virus and the LacZ reporter gene at the deletion site, and construct a recombinant shuttle vector with the LacZ gene as a selection marker; transfect the recombinant shuttle vector into ORFV-infected Vero Homologous recombination was carried out in the cells, and the recombinant virus ORFV-ΔVIR-LacZ with deletion of the VIR virulence gene was obtained by plaque screening and purification. The invention quickly reduces the virulence of ORFV by means of genetic engineering, provides technical support for the research and development of ORF vaccines, has the characteristics of short production cycle, no need for domestication, convenient and quick operation, etc., and can be widely used.
Owner:SHIHEZI UNIVERSITY
Escherichia coli-clostridium shuttle expression vector, and construction and expression thereof
InactiveCN103898098AMicrobiological testing/measurementVector-based foreign material introductionChemical synthesisEscherichia coli
The invention discloses an escherichia coli-clostridium shuttle expression vector, and construction and expression thereof. The escherichia coli-clostridium shuttle expression vector is constructed by inserting escherichia coli lacZ gene (EclacZS) coding sequence between BamHI and SacI restriction enzyme cutting sites of pYN7443 plasmid, wherein the escherichia coli lacZ gene (EclacZS) coding sequence is obtained via clostridium preferred codon chemical synthesis. It is confirmed by escherichia coli lacZ gene activity determination on clostridium transformed from the escherichia coli-clostridium shuttle expression vector that the lacZ gene, which comes from escherichia coli and is obtained via clostridium preferred codon chemical synthesis, is a report gene suitable for expresion in clostridium. Construction of high butanol yield engineered strains via determination of optimal promoter-terminator combination possesses important theoretical significance, wherein determination of the optimal promoter-terminator combination is realized by comparing different promoters and terminators.
Owner:SHANGHAI ACAD OF AGRI SCI
A recombinant fowlpox virus transfer vector expressing chicken type 4 adenovirus fiber2 gene and its construction method and application
ActiveCN107475296BEliminate conflictImprove efficiencyViral antigen ingredientsVirus peptidesTransfer vectorGenetic engineering
Owner:WENS FOODSTUFF GRP CO LTD
HIV drug resistance detection carrier and construction method
ActiveCN109722465AHigh-throughput detectionMicrobiological testing/measurementVector-based foreign material introductionReverse transcriptaseSimian Vacuolating Virus 40
The invention relates to an HIV drug resistance detection carrier, and belongs to the technical field of genetic engineering. A construction method comprises the steps of performing first In-fusion connection on carriers pLWJ-SV40-Luc+ and LacZ genes so as to obtain a carrier pLWJ-SV40-Luc-LacZ, and performing second In-fusion connection on carriers pLWJ-SV40-Luc-LacZ and drug resistance detectiongenes so as to obtain an HIV drug resistance detection carrier, wherein the drug resistance detection genes are genes between 1678bp and 5042bp in an HIV virus Gag-Pol region in a sample to be detected. The HIV drug resistance detection carrier provided by the invention can detect the genes between the 1678bp and the 5042bp in the Gag-Pol region, including HIV-1 reverse transcriptase, protease and integrase inhibitors at the same time, and can cover main action target points of most majority of AIDS antivirus treatment medicines at present, and high-flux detection is realized.
Owner:FUJIAN CENT FOR DISEASE CONTROL & PREVENTION
Kit, specific primer and probe for detecting content of escherichia coli, and application
InactiveCN109055587AAccurate quantification of E. coli contentAccurate quantitative contentMicrobiological testing/measurementDNA/RNA fragmentationEscherichia coliClip seq
The invention provides a group of escherichia coli specific primers and probe sequences so as to establish a fluorescent quantitation PCR detection method which can be applied to detection of the content of escherichia coli in heart blood a dead rat and which is quick, sensitive and good in specificity. According to one side of the invention, specific primers and probes for detecting the escherichia coli are provided, and the specific primers and the probes use conservative fragment sequences as a target goal. A gene clone technique is adopted, escherichia coli LacZ gene specificity fragmentsare inserted into a carrier pMD18-T, so that recombinant plasmids containing LacZ gene specificity fragments are obtained and are used as standard products. A group of specific primers and probes aredesigned and synthesized according to escherichia coli LacZ gene specificity fragment sequences, the PCR reaction condition is optimized, a detection method by using a real-time fluorescent quantitation polymerase chain reaction as a platform is established, and the established method is evaluated.
Owner:上海公安学院 +2
Solvent for chromogenic substrate solution
ActiveUS20060172369A1Material analysis by observing effect on chemical indicatorMicrobiological testing/measurementChromogenic SubstratesSolvent
The present invention relates to a non-toxic dipolar solvent for chromogenic substrate for detecting presence of lacZ gene and / or gene activity, which comprises a stabilizing amount of a solubilizing agent. The present invention also relates to a method for inducing lac operon in screening assay, comprising the step of contacting an agar plate with at least one essential oil in a concentration sufficient to induce the lac operon. The present invention further relates to a method for detecting the presence of bacteria, comprising the step of contacting an agar plate with at least one essential oil in a concentration sufficient to induce detection of the bacteria.
Owner:FISHER SCI CO LLC
Application of high-throughput screening tool for enabling Escherichia coli to obtain effective NHEJ system in Escherichia coli gene editing
PendingCN114277047APromotes non-homologous end joiningImprove efficiencyBacteriaMicrobiological testing/measurementEscherichia coliEnzyme Gene
The invention provides application of a high-throughput screening tool for enabling escherichia coli to obtain an effective NHEJ system in escherichia coli gene editing. The high-throughput screening tool for enabling the Escherichia coli to obtain the effective NHEJ system comprises a pDual-Cas9-Parental plasmid vector and a high-throughput screening vector, wherein the pDual-Cas9-Parental plasmid vector contains a DNA helicase gene, a replicon, an antibiotic resistance gene, a nuclease gene, an araC gene, an arabinose promoter and an IIs type restriction enzyme recognition site; and the pDual-sgRNA-lacZ plasmid vector contains an sgRNA sequence of a targeted lacZ gene, a strong promoter for constitutive expression, a replicon and an antibiotic resistance gene. The NHEJ system obtained through screening has good connection efficiency in escherichia coli, efficient gene editing can be carried out, and the application prospect is wide.
Owner:GENEWIZ INC SZ
Solvent for chromogenic substrate solution
ActiveUS8501433B2Material analysis by observing effect on chemical indicatorMicrobiological testing/measurementChromogenic SubstratesSolvent
Owner:FISHER SCI CO LLC
New lactic acid bacteria
The invention relates to a polynucleotide comprising a lacZ gene (lacZ FS ) encoding a [beta]-galactosidase characterized by a particular profile regarding its efficiency of hydrolysis of lactose. The invention is also directed to a Streptococcus thermophilus strain comprising a lacZ FS allele and bacterial composition thereof, and their use to obtain fermented milk not undergoing post-acidification.
Owner:DUPONT NUTRITION BIOSCIENCES APS
Fowl pox virus double gene expression vector (pg7.5n)
InactiveCN101220374BPromote cloningReduce vector lengthOrganic active ingredientsGenetic material ingredientsOrigin of replicationNucleotide
The present invention provides a fowlpox virus double-gene expression vector, which has the nucleotide sequence shown in SEQ ID NO.1, including promoter P7.5, LacZ gene, multiple cloning site on the LacZ gene, and rare enzyme Cutting site Not1, fowlpox virus DNA homology arms F11L1 and F11L2, AmpR resistance marker gene, origin of replication (ori). The dual-gene expression vector constructed by the present invention minimizes the length of the vector so that larger foreign genes can be accommodated; the number of available restriction sites is increased to facilitate the cloning of different foreign genes; the insertion site Not1 The promoter is not limited, and the promoter that matches with the exogenous gene can be freely selected, thereby improving the expression efficiency, and the expression efficiency of the exogenous gene can be improved by adopting the method of trans expression.
Owner:SOUTH CHINA AGRI UNIV
A rapid screening system for exogenous gene knock-in Lactococcus lactis and its construction method and application
InactiveCN105349565BReduce workloadShorten the timeBacteriaMicrobiological testing/measurementLactococcus lactisBiology
The invention provides a rapid screening system for Lactococcus lactis with knocked-in exogenous genes. The screening system comprises temperature-sensitive plasmids and the Lactococcus lactis, wherein the temperature-sensitive plasmids contain his gene segments and temperature-sensitive replicon-erythromycin resistance gene (Ts-Emr) segments; chromosomes of the Lactococcus lactis contain lacZ gene expression segments. lacZ genes of the Lactococcus lactis cannot be expressed when the exogenous genes are knocked in successfully, so that white colonies are shown on a solid medium containing X-Gal, otherwise, blue colonies are shown, the change from the blue colonies to the white colonies can be directly observed on the solid medium, the screening workload can be greatly reduced, and screening time can be greatly shortened.
Owner:ARMY MEDICAL UNIV
Non-toxic solvent for chromogenic substrate solution and uses thereof
InactiveUS20130316330A1Material analysis by observing effect on chemical indicatorMicrobiological testing/measurementChromogenic SubstratesSolvent
The present invention relates to a non-toxic dipolar solvent for chromogenic substrate for detecting presence of lacZ gene and / or gene activity, which comprises a stabilizing amount of a solubilizing agent. The present invention also relates to a method for inducing lac operon in screening assay, comprising the step of contacting an agar plate with at least one essential oil in a concentration sufficient to induce the lac operon. The present invention further relates to a method for detecting the presence of bacteria, comprising the step of contacting an agar plate with at least one essential oil in a concentration sufficient to induce detection of the bacteria.
Owner:FISHER SCI CO LLC
Method of determining susceptibility of a tumor cell to a chemotherapeutic agent: novel use of herpes
InactiveUS20090325146A1Broaden applicationRapid and non-invasive and reliable assayMicrobiological testing/measurementBiological material analysisApoptosisWilms' tumor
The present invention provides a method of determining if a tumor cell is susceptible to killing by a chemotherapeutic agent, comprising: (a) providing a tumor cell; (b) infecting said tumor cell with a herpes simplex virus or a herpes simplex virus defective in an immediate early gene selected from the group consisting of ICP27, ICP4, and ICP22; and (c) determining the presence of apoptotic killing of said tumor cell, wherein the presence of apoptotic killing is indicative of susceptibility to said chemotherapeutic agent. Chemotherapeutic agent may include doxorubicin, etoposide, paclitaxel, cisplatin, or 5-fluorouracil. The present invention also provides a herpes simplex virus promoter construct having a lacZ gene to assess tumor resistance to chemotherapeutic agents.
Owner:MEDICAL DIAGNOSTIC LAB
A method for co-transformation of oligonucleotide and elimination plasmid for essential gene point mutation of Escherichia coli
The invention relates to a method for carrying out point mutations on essential genes on escherichia coli genomes by using the co-transformation of oligonucleotide and eliminable plasmids. Specifically, front four basic groups express recombinase in escherichia coli by using the co-transformation of oligonucleotide connected by sulfur phosphoryl, with a length of 91 basic groups, and having a mutation site in the middle, and eliminable plasmids. The eliminable plasmids contain homologous sequences of 50 bp lacZ genes, ampicillin resistance genes, I-SceI genes, two I-SceI cleavage sites and R6K replicators. In strains which are integrated with a 50 bp lacZ homologous fragment of a genome due to the catalysis of recombinase so as to express the amicillin resistance, the strains with point mutations are screened. Integrated strains can be induced to express I-SceI under the action of chlortetracycline subjected to heat inactivation, the I-SceI acts on cleavage sites thereof, and through the homologous recombination of two 50 bp lacZ fragments, the eliminable plasmids are eliminated.
Owner:江苏沃纳生物科技有限公司
A recombinant fowl pox virus transfer vector expressing duck type 2 adenovirus fiber2 gene and its construction method and application
ActiveCN107475297BAvoid conflict situationsImprove build efficiencyViral antigen ingredientsVirus peptidesVector vaccineTransfer vector
The invention belongs to the technical field of biology, and mainly relates to a recombinant fowlpox virus transfer vector expressing a duck adenovirus serotype-2 (DADV2) fiber2 gene, a constructing method thereof and applications of the transfer vector. According to the transfer vector, a DADV2 fiber2 gene promoted by a fowlpox-virus-containing early-late promoter LP2EP2, a lacz gene promoted by a P11 promoter, and fowlpox virus genome replicated non-essential fragments which are LTYB and RTYB used for homologous recombination are inserted in a TA cloning site of a pMD19T-Simple vector. The method includes constructing a plasmid pMD-TYB; constructing a plasmid pMD22; constructing a plasmid pMD22-lacz; constructing an intermediate vector pMD22-TYB-lacz; constructing the transfer vector pMD22-TYB-lacz-DFB2; and subjecting the transfer vector pMD22-TYB-lacz-DFB2 to effect verification. The recombinant fowlpox virus transfer vector constructed by the method lays a foundation for development of an efficient recombinant fowlpox virus genetic engineering living-vector vaccine expressing the duck adenovirus serotype-2.
Owner:WENS FOODSTUFF GRP CO LTD
High-throughput screening tool for enabling escherichia coli to obtain effective NHEJ system and application of high-throughput screening tool
PendingCN114164225AShort timeImprove screening efficiencyHydrolasesStable introduction of DNAEscherichia coliEnzyme Gene
The invention provides a high-throughput screening tool for enabling escherichia coli to obtain an effective NHEJ system and an application of the high-throughput screening tool. The high-throughput screening tool comprises a pDual-Cas9-Parental plasmid vector, a plasmid vector and a screening vector, wherein the pDual-Cas9-Parental plasmid vector contains a DNA helicase gene, a replicon, an antibiotic resistance gene, a nuclease gene, an araC gene, an arabinose promoter and an IIs type restriction enzyme recognition site; and the pDual-sgRNA-lacZ plasmid vector contains an sgRNA sequence of a targeted lacZ gene, a strong promoter for constitutive expression, a replicon and an antibiotic resistance gene. The high-throughput screening tool can perform rapid and high-throughput screening on effective Ku + ligD combinations in escherichia coli, multiple Ku + ligD combination results can be obtained through one screening experiment, the time is short, the screening efficiency is high, and conditions are created for gene editing research of escherichia coli.
Owner:GENEWIZ INC SZ
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