39 results about "Promoter mutation" patented technology

The invention discloses a microlunatus phosphovorus engineering strain capable of efficiently accumulating polyphosphate. The microlunatus phosphovorus engineering strain is microlunatus phosphovorus JN459-M571 and is preserved in the China General Microbiological Culture Collection Center with the preservation number of CGMCC No. 11770 on December 1, 2015. A continuous error-prone PCR method is adopted to mutate a promoter of a ppk2 gene in vitro, JN459 is used as a starting strain and a strain library containing the mutant of the promoter of the ppk2 gene is established through a gene recombination technology, and the JN459-M571 strain, of which, the capacity of decomposing polyphosphate under anaerobic conditions is reduced and the capacity of synthesizing polyphosphate under aerobic conditions has no significant difference, is obtained through screening. The Poly-P (polyphosphate) content of the strain disclosed by the invention is 2.4 times higher than that of an original strain JN459 in an anaerobic operation process and the strain disclosed by the invention can be applied to an EBPR (Enhanced biological phosphorus removal) process in a sewage treatment plant.

The invention discloses a probe method detecting human TERT gene promoter mutation and a reagent kit thereof. The method includes the following steps that firstly, human genome DNA is added in a TERT C228T PCR reaction mixed solution so that a C228T reaction system can be formed and fluorescence quantitative PCR amplification can be performed, and / or human genome DNA is added in a TERT C250T PCR reaction mixed solution so that a C250T reaction system can be formed and fluorescence quantitative PCR amplification can be performed; secondly, a result is judged, wherein analysis is performed according to the mutation Ct value of PCR amplification. The reagent kit comprises the TERT C228T PCR reaction mixed solution and / or the TERT C250T PCR reaction mixed solution. The probe method and the reagent kit provide a high-speed, high-sensitivity and high-specificity TERT gene promoter mutation detection system clinically.

The invention belongs to a clinical medicine gene detection technique, and particularly relates to a kit for detecting TERT gene promoter mutation, and a detection method and application of the kit. The kit comprises a fluorescence labeled probe for the TERT gene. Only a pair of probes and a pair of primers are needed in the design, and the detected mutation is specific C228T mutation and accounts for 99% of the bladder cancer TERT mutation. The detection method has the advantages of simple reagents and low price; and the reaction system only contains the most basic primers and probes, so the influencing factors are fewer. The method is accurate for detection and simple to operate, and lays a solid foundation for quick detection of bladder cancer in clinical medicine.

The invention discloses a kit for detecting TERT (telomerase reverse transcriptase) gene mutation, and a detection method of the kit. The kit comprises an LNA (locked nucleic acid) modified specific probe for a TERT gene mutation site, wherein the specific probe can be combined with wild DNA (deoxyribonucleic acid), and can detect a sample containing 0.01% TERT gene mutation DNA; and a minimum detection limit is 2-5cp. The detection method for detecting the TERT gene mutation has the advantages of high specificity, high sensitivity, low pollution, simplicity and rapidness in operation, safety and the like, is especially suitable for detecting the gene mutation from body fluids containing low mutation content such as plasma, urine and saliva, is suitable for performing early screening and diagnosis on a bladder cancer, and provides guidance for personalized medicine.

The invention provides a method for carrying out saturated mutation on known promoters to screen a large number of promoter mutants with different starting activity intensities, and adding modified promoters in front of genes related to amino acid synthesis pathways in industrial amino acid-producing strains to screen strains with further improved amino acid yield, and a mutant promoter sequence obtained from the method and a recombinant strain obtained from the method.

The invention discloses a method and a reagent kit for detecting mutation of a human TERT gene promoter. The reagent kit includes a TERT C228T reaction mixed solution and / or a TERT C250T reaction mixed solution, and further includes a TERT internal standard reaction mixed solution. The method comprises the following steps that firstly, human genome DNA which is extracted from a sample and serves as a template is added into the TERT C228T reaction mixed solution and / or the TERT C250T reaction mixed solution, the human genome DNA same with that in the system is added into an internal standard reaction system, and fluorogenic quantitative PCR amplification is conducted; secondly, Ct values are analyzed. According to the detecting method and the reagent kit, influences of adverse conditions that the GC content of a TERT gene promoter sequence is high and amplification is difficult are overcome, C228T locus mutation and C250T locus mutation are effectively identified, and the detecting sensitivity reaches 1%.

The present invention relates to the field of cancer. More specifically, the present invention provides methods and compositions related to certain promoter mutations in cancer. In one embodiment, a method for treating a subject having thyroid cancer comprises the steps of (a) obtaining a biological sample from the subject; (b) performing an assay on the sample obtained from the subject to identify a mutation at 1 295 228 C>T (C228T) and 1 295 250 C>T (C250T), corresponding to −124 C>T and −146 C>T from the translation start site in the promoter of the telomerase reverse transcriptase (TERT) gene; (c) identifying the subject as having or likely to develop aggressive thyroid cancer if the C228T and / or C250T mutation is identified; and (d) treating the subject with one or more treatment modalities appropriate for a subject having or likely to develop aggressive thyroid cancer.

The invention belongs to the technology of clinical medicine gene detection and particularly relates to a kit for detecting mutation of a TERT gene promoter and a detection method and application thereof. The kit comprises a fluorescence labeling probe specifically recognizing mutation of a lotus C228T and mutation of a lotus C250T or other combinations of the TERT gene promoter, a pair of TERT specific primers and needed Master mix of all reagents. The specificity mutation C228T and the specificity mutation C250T of the TERT gene promoter are provided, and the sum of the specificity mutation C228T and the specificity mutation C250T accounts for about 100% of bladder cancer TERT mutation; a detection method is standardized, the reagents are subjected to optimization verification, the price is low, all ingredients in a reaction system are premixed, the operation process is simplified to the maximum extent, and operation errors are avoided. When the kit is used for detecting mutation of the TERT gene promoter, the number of influence factors is small, detection is precise, operation is easy, detection results are understood easily, and a reliable method for quickly detecting bladder cancer through urine is provided for clinical medicine.

The present invention relates to the field of cancer. More specifically, the present invention provides methods and compositions related to certain mutations in cancer. In one embodiment, a method for treating a subject having aggressive thyroid cancer comprises the steps of (a) obtaining a biological sample from the subject; (b) performing an assay on the sample obtained from the subject to identify a mutation at 1 295 228 C>T (C228T), corresponding to −124 C>T from the translation start site in the promoter of the telomerase reverse transcriptase (TERT) gene, and a T1799A mutation in the BRAF gene that results in a V600E amino acid change; (c) identifying the subject as having or likely to develop aggressive thyroid cancer if the C228T and V600E mutations are identified; and (d) treating the subject with one or more treatment modalities appropriate for a subject having or likely to develop aggressive thyroid cancer. Similar approaches are applied to other human cancers harboring both BRAF V600E mutation and TERT promoter mutations.

The invention discloses a core promotor sequence for regulating expression of a plutella xylostella Bt (Bacillus Thuringiensis) Cry1Ac insecticidal protein resistant gene MAP4K4 (Mitogen-activated Protein Kinase Kinase Kinase Kinase 4), mutation sites and application of the core promotor sequence in plutella xylostella Bt Cry1Ac insecticidal protein resistance identification and field monitoring.The mutation sites are nucleotides located at a 491 site, a 474 site and 469 to 470 sites at the upstream of an initial codon ATG and are respectively named M1, M2 and M3. A nucleotide sequence of plutella xylostella Bt Cry1Ac insecticidal protein susceptible population promotor is as shown in SEQ ID NO.1, and a nucleotide sequence of a resistance near-isogenic line population promotor is as shownin SEQ ID NO.2. An experiment verifies that the activity of a MAP4K4 gene promotor is remarkably enhanced when M2 and M3 are in mutation at the same time. According to the core promotor sequence disclosed by the invention, the research of promotor mutation in the field of insect Bt resistance is promoted, and particularly, important theoretical and practical significances in aspects of insect Btresistance field detection and control are obtained.

The present invention relates to the field of cancer. More specifically, the present invention provides methods and compositions related to certain mutations in cancer.In one embodiment, a method for treating a subject having aggressive thyroid cancer comprises the steps of (a) obtaining a biological sample from the subject; (b) performing an assay on the sample obtained from the subject to identify a mutation at 1 295 228 C>T (C228T), corresponding to -124 C>T from the translation start site in the promoter of the telomerase reverse transcriptase (TERT) gene, and a T1799A mutation in the BRAF gene that results in a V600E amino acid change; (c) identifying the subject as having or likely to develop aggressive thyroid cancer if the C228T and V600E mutations are identified; and (d) treating the subject with one or more treatment modalities appropriate for a subject having or likely to develop aggressive thyroid cancer. Similar approaches are applied to other human cancers harboring both BRAF V600E mutation and TERT promoter mutations.

The invention uses Bacillus as a starting strain, uses a carbon metabolism response element cre in a maltose promoter to transform a target, a lysine ribose switch is fused at the same time, a mutantof the maltose promoter is constructed, the mutant that can reduce the leaky expression of the maltose promoter and improve the induced expression strength of the maltose promoter is obtained, and anexpression vector and host cell which contain the mutant of the promoter are constructed, so as to be conducive to the application of the maltose promoter in protein expression.

The invention discloses a method, a primer and a kit for detecting mutation sites of promoters C228T and C250T of a telomerase reverse transcriptase (TERT) gene. The primer comprises a forward primer and a reverse primer for amplifying the mutation sites of the promoters C228T and C250T of the TERT gene, and in addition, can also comprise one pair of sequencing primers. A Touch-down PCR (Polymerase Chain Reaction) amplification technique is combined with a Sanger sequencing method; the occurrence conditions of the mutation sites of the promoters C228T and C250T of the TERT gene in a body of a patient suffered from a brain glioma can be quickly detected.

The invention relates to a Trc promoter mutation library and application thereof, and belongs to the technical field of biology. Promoters shown in SEQ ID NO.1-41 are obtained through a large number of screening, and the range of regulating and controlling the intensity of a target gene table is wide so that a Trc promoter mutation library is established. According to the invention, the promoter library is constructed, and the opening amount type fine adjustment gene expression becomes possible; in metabolic analysis and transformation, the disturbance on the cell genotype can be realized onlyby two non-'on ', namely'off' means of gene knockout and overexpression in the past, namely, the influence of the target gene expression on phenotype and metabolic flux distribution can be analyzed and controlled by performing gradient type accurate fine adjustment on the target gene expression.

The invention discloses an NGS-based brain tumor molecular diagnosis analysis method. The method comprises the following steps: constructing a cluster based on an Amazon cloud platform; establishing an analysis process of the molecular marker for analyzing the brain tumor on the cluster; a sequence alignment module; removing the repetitive sequence module; a gene fusion analysis module; a call variation module; a cnv analysis structure variation module; the method disclosed by the invention has the advantages of high sensitivity, high specificity, high repeatability, low experimental cost andshort analysis period, and can be used for comprehensively analyzing the hot spot mutation of the TERT promoter, the deletion states of the chromosomes 1p and 19q and other main brain tumor related molecular markers at one time. The NGS-based analytical technique of the present invention enables reliable detection of 1p and / or 19q deletions in the background tumor tissue of 70% normal cells, is more sensitive than microsatellite-based LOH analysis, and requires less DNA.

The invention provides a method for quickly constructing and screening a maltose transcriptional activation factor MalR mutation library through fluorescent protein by using bacillus as an original strain, and besides, through screening, a promoter mutant which can reduce leakage expression of the maltose promoter and increase the induction expression intensity is obtained by screening, so that anapplication of the maltose promoter to protein expression is facilitated.

The invention relates to the technical field of detection reagents or kits for coronary heart disease susceptive genes and discloses a reagent for in-vitro detection on mutation of an SelS gene promoter and application of the reagent in preparation of a coronary heart disease screening kit. A nucleotide sequence of the SelS gene promoter is represented by SEQ ID NO: 1 of a sequence listing, and when the SelS gene promoter is mutated, loci 84 to 89 of the nucleotide sequence are deleted. The invention discloses a novel SelS gene promoter mutant site closely related to susceptivity of a coronaryheart disease, and a kit for screening or predicting the coronary heart disease is developed based on the promoter mutant site. According to the application of the SelS gene promoter mutant site disclosed by the invention, early screening of the coronary heart disease can be more accurately achieved from a molecular level, the pertinence is high, and the sensitivity is high.

The invention discloses a kit for detecting TERT (telomerase reverse transcriptase) gene mutation, and a detection method of the kit. The kit comprises an LNA (locked nucleic acid) modified specific probe for a TERT gene mutation site, wherein the specific probe can be combined with wild DNA (deoxyribonucleic acid), and can detect a sample containing 0.01% TERT gene mutation DNA; and a minimum detection limit is 2-5cp. The detection method for detecting the TERT gene mutation has the advantages of high specificity, high sensitivity, low pollution, simplicity and rapidness in operation, safety and the like, is especially suitable for detecting the gene mutation from body fluids containing low mutation content such as plasma, urine and saliva, is suitable for performing early screening and diagnosis on a bladder cancer, and provides guidance for personalized medicine.

The invention relates to methods and kits for identifying individuals at risk of thiopurine drug intolerance based on detecting the presence of mutations in the TPMT gene promoter associated with thiopurine drug resistance or intolerance.

The invention discloses a method for screening and replacing a pET commercial plasmid promoter. According to the method, heterologous expression of nitrilase is adjusted from the transcriptional level, the correct folding proportion of nitrilase is increased, soluble expression is increased, and inhibition of inclusion body accumulation on thallus growth is relieved; the promoter mutant strain is subjected to shake-flask induced expression, and the thallus concentration, the specific enzyme activity and the fermentation liquor specific enzyme activity are 1.4 times, 1.6 times and 5.5 times higher than those of commercial plasmids respectively; meanwhile, in the reaction of catalytically preparing the gabapentin intermediate 1-cyanocyclohexyl acetic acid; and the catalytic efficiency of the promoter mutant strain is improved by more than 100% compared with that of the original strain.

The present disclosure provide for methods of using 6-thio-2′-deoxyguanosine (6-thio-dG) to treat telomerase-positive cancers that exhibit (a) one or more TERT promoter mutations, and / or (b) enriched telomere transcriptional signature(s). In particular, melanomas, including those who are not sensitive or have become resistant to immune checkpoint inhibition and / or MAPKi therapy are targets for this therapy.

The invention provides a primer composition, kit and method for detecting TERT gene promoter mutation and application of the primer composition. The primer composition comprises three pairs of TERT primers of SEQ ID NO: 1 to SEQ ID NO: 6; and SEQ ID NOs: 4 to 6 respectively have different barcode sequences. The primer composition furthermore comprises a general primer and three reverse primers with different indexes. By designing forward and reverse primer sequences covering the CTBT and C250T mutation sites of a TERT gene promoter, an amplification product can include the mutation sites C228Tand C250T and all other potential mutation sites. By performing a second round of PCR amplification on the amplification product of the above primers, a linker sequence is complemented, and the abovemutation is directly detected by utilizing a high-throughput sequencing method after purification, so that the advantages of being simple to operate, high in sensitivity and low in cost are provided.

The present application discloses Zymomonas mobilis endogenous promoter mutants which guide the expression of heterologous nucleic acids that can be chimeric-linked, promote the expression of heterologous nucleic acids, and at the same time, these promoter mutants are high-expression promoters having a significantly enhanced function relative to wild-type promoters, the present invention relates to chimeric genes for expressing chimeric genes in cells of Zymomonas and / or Escherichia.

The invention relates to an internal reference plasmid suitable for a silkworm cell double luciferase reporter gene system and a preparation method and application of the internal reference plasmid. The internal reference plasmid contains a constitutive promoter BmVgP78M suitable for the silkworm cell double luciferase reporter gene system, wherein the promoter is obtained by a BmVg gene promoter mutant hormone element to constitute the internal reference plasmid which does not respond to a silkworm hormone, can be stably expressed in silkworm cells, has moderate activity, and can be effectively used for the silkworm cell double luciferase reporter gene system to study the promoter activity or gene regulation.

The present invention relates to mutations in the amyloid precursor protein (APP) promoter region, whereby the mutations cause a significant increase in APP expression. The increase in APP expression is related to Alzheimer's disease, and the mutations can be used in Alzheimer's disease diagnosis, or in the construction of transgenic animal models for studying Alzheimer's disease.

The invention provides a mouse triple-negative primary glioblastoma cell strain G422TN-GBM, which is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: C2020267. The mouse triple-negative primary glioblastoma cell strain provided by the invention has the following molecular characteristics: an IDH1 / 2 wild type is adopted, Chr1 / 19 is complete, TERT promoter mutation is negative, and ATRX mutation and Trp53 mutation are accompanied. A glioblastoma animal model established by the cell strain has high replicability and stability, can simulate the clinical treatment response of glioblastoma at each stage, can be used for anti-tumor drug screening, new therapy pre-clinical curative effect evaluation and single-drug and combined treatment scheme curative effect evaluation, and has a wide application prospect. And evaluating the early, middle and late treatment effects of the malignant glioma.

The invention discloses a Trichoderma reesei cbh1 gene promoter mutant and its construction method and application, belonging to the technical field of genetic engineering; the Trichoderma reesei cbh1 gene promoter mutant provided by the invention has a nucleotide sequence such as SEQ ID No .2 shown. The present invention also provides a recombinant strain of Trichoderma reesei, which contains the above-mentioned Trichoderma reesei cbh1 gene promoter mutant. The Trichoderma reesei cbh1 gene promoter mutant constructed by the present invention can improve the effect of driving the expression of β-mannanase, and the expression of the β-mannanase constructed by the Trichoderma reesei cbh1 gene promoter mutant of the present invention can be improved The strain can significantly increase the production of β-mannanase.

The present invention provides a method for rapidly constructing and screening maltose transcription activator MalR mutation library using fluorescent protein as the starting strain, and at the same time screens to obtain a promoter mutation that can reduce the leaky expression of the maltose promoter and increase its induced expression intensity body, which is beneficial to the application of maltose promoter in protein expression.

The invention discloses a core promotor sequence for regulating expression of a plutella xylostella Bt (Bacillus Thuringiensis) Cry1Ac insecticidal protein resistant gene MAP4K4 (Mitogen-activated Protein Kinase Kinase Kinase Kinase 4), mutation sites and application of the core promotor sequence in plutella xylostella Bt Cry1Ac insecticidal protein resistance identification and field monitoring.The mutation sites are nucleotides located at a 491 site, a 474 site and 469 to 470 sites at the upstream of an initial codon ATG and are respectively named M1, M2 and M3. A nucleotide sequence of plutella xylostella Bt Cry1Ac insecticidal protein susceptible population promotor is as shown in SEQ ID NO.1, and a nucleotide sequence of a resistance near-isogenic line population promotor is as shownin SEQ ID NO.2. An experiment verifies that the activity of a MAP4K4 gene promotor is remarkably enhanced when M2 and M3 are in mutation at the same time. According to the core promotor sequence disclosed by the invention, the research of promotor mutation in the field of insect Bt resistance is promoted, and particularly, important theoretical and practical significances in aspects of insect Btresistance field detection and control are obtained.