65 results about "ALDH2 gene" patented technology

The invention provides an sg RNA sequence used for knocking out a human ALDH2 gene;,a target DNA sequence of the sg RNA is at least one of sequences as shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3. The invention further provides a method for knocking out a human hepatoma carcinoma cell ALDH2 gene; the method utilizes a CRISPR / Cas system to modify the ALDH2 gene in a human hepatoma carcinoma cell. The invention further provides two ALDH2 gene deletion cell strains; ALDH2 participates in an important metabolic function of a body. The ALDH2 gene deletion cell strains provided by the invention provide an effective platform for metabolism study, in vivo, of exogenous chemicals or exogenous poisons, so that powerful means are provided for researching chronic diseases (such as alcoholic liver diseases and diabetes) as well as tumor-associated diseases.

The invention provides a human ALDH2 genotype detection kit comprising a PCR amplification primer, DNA polymerase and a reaction buffer solution; wherein the DNA polymerase is a polyethylene glycol (PEG) modified hot-start Taq polymerase which can tolerate the PCR reaction inhibitors in blood; and further provides the nucleotide sequence of the PCR amplification premier. The kit has the following advantages: (1) the DNA polymerase is a polyethylene glycol (PEG) modified hot-start Taq polymerase which can tolerate the PCR reaction inhibitors in blood, thus can realize the human whole-blood PCR amplification without carrying out nucleic acid extraction, and is capable of directly carrying out PCR amplification in blood; (2) the kit can carry out detection on a sample ALDH2 genotype.

The invention discloses a kit for detecting alcoholic liver disease susceptibility. The kit detects three genes closely related with an alcoholic liver disease, namely an ADH2 gene of alcohol dehydrogenase, an ALDH2 gene of acetaldehyde dehydrogenase and a CYP2E1 gene of cytochrome P4502E1. The simultaneously detected SNP sites comprise an rs1229984 site of the ADH2 gene, an rs671 site of the ALDH2 and an rs2031920 site of the CYP2E1. Whether a detected crowd carries ''alcoholic liver disease susceptible genes'' can be clearly judged by using the kit, detecting a group of genes and sites related with the alcoholic liver disease susceptibility, using specific primers and probes and combining mononucleotide extension technology and micro array chip technology, and the alcoholic liver disease susceptible crowd is screened from the crowd, so that poor life habits are changed and the purpose of prevention is achieved.

The invention discloses a genetically engineered bacterium for expressing a human derived acetaldehyde dehydrogenase gene and application thereof, and belongs to the technical field of gene engineering and enzyme. The ALDH2 gene efficient expression component Trp1L-URA3-TPIp-ALDH2-TPIt-Trp1R is integrated into the genome of Saccharomyces cerevisiae W303-1A, the stability of gene expression is improved compared with free plasmid expression, a constitutive promoter is used, no induction agent is added during fermentation, and the product is relatively safe. When fermentation is performed for 96hours, the specific activity of producing acetaldehyde dehydrogenase reaches 0.97 U / mg. The specific activity of the producing acetaldehyde dehydrogenase reaches 6.256 U / mg in ethanol with the concentration of 250 mg / 100 mL, and the specific activity of the producing aldehyde dehydrogenase in a buffer with pH of 3 reaches 0.301 U / mg. The Saccharomyces cerevisiae is selected as a host strain, on the one hand, the activity of the enzyme is kept, on the other hand, product safety is improved, and a new approach and a theoretical basis are provided for the research and development of antialcoholicdrugs.

The invention belongs to the field of biotechnology and relates to a kit for typing genes related to alcohol, folic acid and VD metabolic capability of a human body based on a SNaPshot technique. The kit can be used for once quickly detecting the gene types of ADH1B gene G1434A locus, ALDH2 gene G1510A locus, MTHFR gene A1298C locus and C677T locus, MTRR gene A66G locus and VDR gene Bsm I, Apa I and Fok I locus. The kit comprises a PCR amplification reagent, a PCR product purifying reagent, an extending reaction reagent, an extension reaction purifying reagent and an electrophoresis reagent, wherein the PCR amplification reagent comprises an amplimer mixed solution I of 8 locus of 5 related genes and the extension reaction purifying reagent comprises an amplimer mixed solution II of 8 locus of 5 related genes. The kit provided by the invention can once quickly, accurately, sensitively and economically detect the related gene type of human metabolic capability including alcohol metabolic capability, folic acid utilizing capability and vitamin D utilizing capability and can supply scientific and reliable basis to human health management.

The invention relates to a sequencing primer pair for detecting an ALDH2 (Aldehyde Dehydrogenase 2) genotype with a pyrosequencing method and a kit thereof, and belongs to the technical field of in-vitro nucleic acid detection. The primer pair comprises a positive amplification primer, a reverse amplification primer and a sequencing primer, wherein a 5' end of the positive amplifier primer is subjected to biotin labeling. The kit comprises the positive amplification primer, PCR (Polymerase Chain Reaction) reaction liquid containing the reverse amplification primer, the sequencing primer, uracil DNA (Deoxyribose Nucleic Acid) glycosylase and Taq polymerase. The kit provided by the invention has the advantages of accurate detection result, high specificity, short detection period, easiness in operation, and capability of effectively meeting clinical examination requirements. Moreover, the kit further has the advantages that a reaction process can be monitored in real time, the reaction time is short, a PCR product can be subjected to pyrosequencing by a pyrosequencing instrument and high-flux sample detection through simple treatment, and the sensitivity is higher compared with a golden standard method, namely, a capillary electrophoresis sequencing method.

The invention relates to a method and kit for detecting human alcohol metabolizing capacity gene mutation sites. The detection method comprises the following steps: designing specific primers by aiming at an alcohol dehydrogenase ADH1B gene rs1229984 site and an acetaldehyde dehydrogenase ALDH2 gene rs671 site, and performing specific polymerase chain reaction (PCR) amplification so as to obtain target fragments; and then performing enzyme disgestion, single base extension and desalting purification treatment, and then detecting analytic sequences by use of a nucleic acid velocitron. The method for detecting the human alcohol metabolizing capacity gene mutation sites is high in detection accuracy, high in experiment repeatability, great in flux and low in cost. The invention also providesa kit for detecting human alcohol metabolizing capacity gene mutation sites. The kit comprises a specific primer pair for amplifying an alcohol dehydrogenase ADH1B gene rs1229984 site gene and an acetaldehyde dehydrogenase ALDH2 gene rs671 site gene. The kit for detecting human alcohol metabolizing capacity gene mutation sites is capable of simplifying experimental procedures and has the advantages of short manual operation time, low difficulty and high experiment automation degree.

The invention belongs to the technical field of molecular biological gene polymorphism detection, and in particular relates to a kit and a method for genotyping detection. The kit consists of a firstprimer pair and a first molecular beacon probe for ADH1B gene rs1229984 site, a second primer pair and a second molecular beacon probe for ALDH2 gene rs671 site, a third primer pair and a third molecular beacon probe for CYP1A2 gene rs762551 site, and a fourth primer pair and a fourth molecular beacon probe for TMPRSS6 gene rs855791 site. With the application of the kit provided by the invention,four genotypes corresponding to the genes and corresponding metabolic absorption types can be rapidly and conveniently detected.

The invention belongs to the technical field of biology and specifically relates to a human ALDH2 gene polymorphism detection kit as well as a preparation method and application thereof. The kit is prepared from PCR premixed reaction liquid, a positive control product and a negative control product, wherein the PCR premixed reaction liquid is used for amplifying a G1510A site of ALDH2 gene; the PCR premixed reaction liquid is prepared from a specificity primer sequence set, a probe set and PCR reaction liquid; the specificity primer sequence set is used for amplifying all the sites; the specificity primer set is prepared from an ordinary outer primer and an ARMs primer with fluorescent label specificity. The kit disclosed by the invention is used for detecting the human ALDH2 gene polymorphism and has the advantages of high flexibility, high specificity, convenience in operation, reliable results and the like; furthermore, detection can be finished within 1 hour, and result reading issimple and objective.

The invention discloses a primer and a kit for detecting genetic typing of human aldehyde dehydrogenase 2 (ALDH2). The primer group for detecting human ALDH2 gene typing comprises an ALDH2*1 primer pair, an ALDH2*2 primer pair and an internal reference primer pair. The application of the kit helps to realize good typing for ALDH2 gene, and has reference value for hereditary dangerous factors such as drinking behavior, alcoholic cirrhosis, liver cancer, nasopharyngeal carcinoma, oral cancer and the like, and glyceryl trinitrate nitroglycerin guiding pharmacy.

The invention discloses a kit for detecting acetaldehyde dehydrogenase 2 (ALDH2) gene polymorphism by a pyro-sequencing method and a method. By the kit, the ALDH2 gene polymorphism, particularly RS671 single nucleotide polymorphism can be detected. The kit comprises primers which are shown as SEQ ID NO.2-4. By the kit, the accurate, quick and high-throughout detection of ALDH2 gene polymorphism can be realized, so that the alcohol tolerance can be effectively predicted and prevented, and the clinical individual administration of nitroglycerin can be guided.

The invention relates to the field of molecular biology, and discloses a specific oligonucleotide probe for detecting an SNP (single nucleotide polymorphism) site of ALDH2 (alcohol dehydrogenase 2)gene, wherein the probe is capable of hybridizing with the 504th different genetype of the ALDH2 gene. The invention further discloses a specific primer pair for amplifying the ALDH2 gene when detecting the SNP site, and the primer pair can be used for specifically amplifying a target area containing a detection target site in the ALDH2 gene. The specific oligonucleotide probe and the specific primer pair provided by the invention can be used for detecting the ALDH2 genetype and providing information for people to learn individual alcohol metabolism detoxification ability and correctly use nitroglycerin.

The invention relates to a detecting method for evaluating the curative effect of nitroglycerin and the similar medicament which treat angina pectoris, and the method comprises utilizing a sampling mop to scrap at least 40-60 times from top to bottom on the left and the right cheeks in the mouth of a measured person, gathering enough cell samples, extracting genome DNA, detecting the variation of mitochondria acetaldehyde dehydrogenase gene ALDH2 Glu504Lys, and evaluating the curative effect of nitroglycerin and the similar medicament which treat angina pectoris through detecting and analyzing the Glu504Lys site genotype on an individual ALDH2 gene. The invention has the advantages that the detecting method can be used to evaluate the validity of nitroglycerin and the similar medicament which treat angina pectoris.

The invention discloses an ALDH2 (aldehyde dehydrogenase 2) Gene polymorphism detection kit, having the advantages that detection sensitivity is high, the lowest detection limit is 7.8 pg DNA; the kitis simple to operate and allows point-of-care test without professional knowledge; the detection time is short, down to 45 min; the detection specificity is high, and the detection accuracy is very high since a specific fluorescent probe is used; nucleic acid extraction is not required, low-melting-point agarose is added, and PCR (polymerase chain reaction) is greatly avoided. When used with ParaDNA gene amplification detectors to carry out POCT (point-of-care test), the kit can be used to clinically provide guidance on the administration of nitroglycerin and the notification on the risk of alcohol drinking.

The present invention discloses a kit and a method for detecting ALDH2 polymorphism based on a FRET probe melting curve. The kit comprises a forward primer and a reverse primer for specifically amplifying the gene ALDH2 (rs671) locus, and a pair of fluorescently labeled probes for specifically distinguishing the gene ALDH2 (rs671) polymorphism, wherein the specific sequence of the forward primer is F: 5'-AGCAGACCCTAAATCCCTGG-3, the specific sequence of the reverse primer is R: 5'-CACCAGCAGACCCTCAAG-3', the specific sequence of the sensor probe is 5'-TGCAGGCATACACTGAAGTGAAAACTGT-FAM-3', and the specific sequence of the anchor probe is 5'-ROX-AGTGTGGGACCTGCTGGGGGCT-P-3'. According to the present invention, the advantages of good specificity, high sensitivity, low price, simple operation and avoidance of pollution caused by tune opening operation are provided, and the result can be intuitively and clearly interpreted through the TM value read by the instrument.

The invention discloses a reagent box for detecting alcoholic intoxication and susceptibility to alcohol habituation. The reagent box comprises specificity primer pairs and specificity fluorescent probes for detecting position 161 SNP position in ALDH2 gene SEQ ID NO:1, and an ordinary assembly for fluorescent quantitative PCR detection. The reagent box predicts the alcoholic intoxication and susceptibility to alcohol habituation of an individual by detecting and analyzing the gene carried on position SNP on ALDH2 gene.

The invention belongs to the technical field of biological detection and discloses a kit for accurately detecting the polymorphism of an ALDH2 (Aldehyde Dehydrogenase 2) gene. The kit for accurately detecting the polymorphism of the ALDH2 gene can be used for determining mutation gene types (G>A, G>T and G>C) of rs671 polymorphism sites of the ALDH2 gene in one step. In order to realize the function, the kit applies a method combining a multi-PCR (Polymerase Chain Reaction) technology and a Taqman-MGB fluorescent probe labeling technology, and four pairs of specific primers and four specific probes for labeling different fluorescent dyestuffs are designed; the kit is characterized in that a target gene only can be combined with one pair of primers and one probe, so that the accuracy of a detection result is ensured. The kit provided by the invention has the advantages of strong specificity, high accuracy, simplicity in operation and the like. A sample to be detected can be selected from whole blood or oral cells; the kit is clinically used for guiding rational drug use of nitroglycerin and guiding healthy wine drinking to prevent alcohol intoxication, and has great significance inthe aspect of prompting major diseases.

The invention discloses a reagent kit for detecting polymorphism of a human ALDH2 (acetaldehyde dehydrogenase 2) gene by a TaqMan-MGB (minor groove binder) probe method. The reagent kit comprises the following nucleic acid sequences: (1) a primer pair for amplifying an rs671 polymorphism site of the ALDH2 gene, and (2) a TaqMan-MGB probe for detecting the rs671 polymorphism site of the ALDH2 gene, wherein a nucleotide sequence of the primer pair is shown as SEQ ID NO. 1 and SEQ ID NO. 2; a nucleotide sequence of the TaqMan-MGB probe is shown as SEQ ID NO. 3 and SEQ ID NO. 4; the SEQ ID NO. 3 is used for labeling an FAM signal; and the SEQ ID NO. 4 is used for labeling a VIC signal. The invention further discloses an application of a primer and the probe in detecting the polymorphism of the human ALDH2 gene. The primer has the advantages of accurate detection result, quickness, high efficiency, convenience and high sensitivity.

The invention provides a gene detection reagent kit for evaluating alcohol tolerance. The gene detection reagent kit comprises six amplification primers for three genes including an internal reference gene GAPDH (glyceraldehyde phosphate dehydrogenase), a gene ADH1B (ethanol dehydrogenase 1B) and a gene ALDH2 (acetaldehyde dehydrogenase 2) and five Taq-man probe primers. One of the Taq-man probe primers is positioned at the internal reference gene GAPDH and is used for carrying out FAM fluorescence labeling; two other Taq-man probe primers are SNP (single nucleotide polymorphism) probes positioned at the gene ADH1B and are used for carrying out VIC and ROX (roxithromycin) labeling; the remaining two Taq-man probe primers are SNP probes positioned at the gene ALDH2 and are used for Cy5 and Quasar705 labeling, BHQ is used as a quenching group, and delta ct values can be computed. The gene detection reagent kit has the advantages that the alcohol tolerance of healthy persons can be evaluated by the gene detection reagent kit from the aspect of genes, and accordingly the healthy persons can know the strength and the weakness of alcohol tolerance, can drink alcohol scientifically and can carry on occasion such as party, gathering and banquet related to alcohol culture.

The invention belongs to the technical field of gene engineering and discloses a primer and a probe combination for detection of human ALDH2 gene polymorphism, a kit comprising the primer and the probe combination, and a fluorescent PCR method for performing the detection of human ALDH2 gene polymorphism with the primer and the probe combination. The primer, the probe and the kit are based on TaqMan fluorescent PCR technology, and are simple, quick and high-sensitive. In addition, through reasonable combination of primer and probe, interaction between primer and primer, primer and probe and probe and probe can be avoided effectively, thus reducing detection error. The primer, the probe and the kit, when being used for the detection of human ALDH2 gene polymorphism, have high sensitivity and strong specificity, are easy, quick and safe to operate, have simple and direct results, and can be applied directly with a blood sample or a dried blood spot specimen on filter paper as a template.

The invention provides primers and probes for detecting an ALDH2 (aldehyde dehydrogenase gene), and a detection method for detecting the ALDH2. The detection method is characterized in that primers and probes of heterozygous ALDH2*1 / *2, homozygous ALDH2*2 / *2 and internal reference gene beta-globin are used for preparing PCR (Polymerase Chain Reaction) liquid; human whole blood sample DNA (Deoxyribonucleic Acid) is added; and the polymorphism of the ALDH2 is detected through a fluorescent PCR instrument. According to the method, the specificity detection probe is used for recognizing a target gene, and the accuracy is high. Meanwhile, a target sequence is controlled in a duplex way by the primers and the probes; the specificity is good; the false positive rate is low; the sensitivity is 0.01 percent, i.e., the detection can be realized when one cell with the polymorphism of the ALDH2*1 / *2 of the ALDH2 and the ALDH2*2 / *2 of the ALDH2 exists in 10,000 cells; the amplification and the detection can be performed in the same tube; the cover opening is not needed; the pollution cannot easily occur; meanwhile, the amplification and the detection can be completed in one step; later-stage processing is not needed; and the whole-process monitoring is realized. A kit provided by the embodiment of the invention introduces an internal positive control quality control system; the whole-process quality monitoring is realized in the detection process; the false positive result or the false negative result is effectively avoided; the speed is high; and the process can be completed in 100 minutes.

The invention discloses a probe for detecting mutation of a gene ALDH2, as well as application thereof and a kit, and belongs to the technical field of gene detection. The probe comprises a detection probe as shown by SEQ ID NO.1, and one or two of capture probes as shown by SEQ ID NO.2-3. The probe can detect G to A mutation at site rs671 of the gene ALDH2, and has the characteristics of high detection sensitivity, strong specificity, low false positive rate, and the like.

The invention discloses a detection method of a gene related to alcohol metabolism. The detection method comprises the specific steps that DNA of a detected person is extracted, and the concentration of the extracted DNA is not lower than 10 ng / ul, 260 / 280=1.8; SEQ NO 1 and SEQ NO 2 are utilized to amplify the rs1229984SNP locus of an ADH1B gene, SEQ NO 3 and SEQ NO 4 are utilized to amplify the rs671SNP locus of an ALDH2 gene, SEQ NO 5 and SEQ NO 6 are utilized to amplify the rs3813867 and rs2031920 SNP loci of a CYP2E1 gene, and the extracted DNA serves as a template to conduct PCR reaction; the three pairs of primers are subjected to agarose gel electrophoresis which is 1% of a common PCR amplification product; the 1% agorose gel electrophoresis strip is used for recycling a target strip by using an agarose gel extraction kit; sequencing reaction is conducted; a product obtained through the sequencing reaction is purified, denatured, and subjected to sequencing on a 3730 sequenator; and Chromas software is utilized to analyze the four SNP gene types from the sequencing result. The detection method of the gene related to alcohol metabolism has the advantages of being capable of conveniently conducting detection, high in detection accuracy and the like.

The invention discloses primers and a reagent kit for detecting the polymorphism of the ALDH2 gene c.1510 locus. The specific primers, a specific probe and the specific reagent kit which are designed for the ALDH2 gene locus have the advantages of being high in sensitivity, good in specificity, high in response speed and low in cost, and are suitable for large-scale clinical application; rapid, effective and accurate typing qualitative detection of ALDH2 can be achieved, and a reference can be provided for timely nitroglycerin treatment of stenocardia, drinking guidance and occurrence of related caner.

The invention provides an acetaldehyde dehydrogenase recombinant gene as well as a lactic acid bacteria vector and application thereof. The nucleotide sequence of the recombinant gene is as shown in SEQ ID NO: 1. The amino acid sequence of encoded human-derived acetaldehyde dehydrogenase is shown as SEQ ID NO: 2. The artificial synthesis method of the recombinant gene comprises the following steps: carrying out codon optimization on an ALDH2 gene sequence obtained by an NCBI database to obtain an ALDH2-CO (Codon optimized) enzyme gene; separately introducing restriction endonucleases cutting sites KpnI and EcoNI to the 5'end and the 3 'end of the ALDH2-CO enzyme gene, and performing PCR amplification, agarose gel electrophoresis and gel recovery in sequence to obtain an ALDH2 target gene segment; and connecting the ALDH2 target gene segment to a lactic acid bacteria expression vector, and converting the gene segment into lactic acid bacteria cells. The invention provides the novel recombinant acetaldehyde dehydrogenase gene and lactic acid bacteria expressing the recombinant gene, and provides a new way and theoretical basis for research and development of hangover alleviating products.

A primer set and probe set for predicting alcohol-degrading ability and hangover development is provided to improve rapidness and convenience of prediction by amplifying at least one target sequence selected from ALDH2(acetaldehyde dehydrogenase 2) gene, CYP2E1(cytochrome P450 2E1) gene and ADH2(alcohol dehydrogenase 2) gene. An oligonucleotide probe set capable of hybridizable with at least one target sequence selected from exon XII region of ALDH2 gene, 5'-regulating region of CYP2E1 gene and the exon III region and exon IX region of ADH2 gene is selected from (1) an oligonucleotide probe capable of hybridizable with the exon XII region of ALDH2 gene containing 10 or more consecutive nucleotide fragments containing a 11st base in the nucleotide sequence of SEQ ID NO:15, (2) an oligonucleotide probe capable of hybridizable with the 5-regulating region of CYP2E1 gene containing 10 or more consecutive nucleotide fragments containing a 12nd base in the nucleotide sequence of SEQ ID NO:17, (3) an oligonucleotide probe capable of hybridizable with the exon III region of ADH gene containing 10 or more consecutive nucleotide fragments containing a 12nd base in the nucleotide sequence of SEQ ID NO:19, (4) an oligonucleotide probe capable of hybridizable with the exon IX region of ADH2 gene containing 10 or more consecutive nucleotide fragments containing a 12nd base in the nucleotide sequence of SEQ ID NO:21, and (5) an oligonucleotide probe capable of hybridizable with the exon XII region of ADH2 gene containing 10 or more consecutive nucleotide fragments containing a 13rd base in the nucleotide sequence of SEQ ID NO:23.

A probe for detection of at least 1 type of genetic polymorphism of the ALDH2 gene rs671 and the ADH2 gene rs1229984, a kit therefore, and methods of detecting the polymorphism(s).

The invention discloses a method, primer and detection kit for polymorphism detection of two genes of ethanol dehydrogenase ADH1B and acetaldehyde dehydrogenase ALDH2 which are relative to alcohol metabolism. Both the primer and the detection kit comprise forward and reserve primers amplifying polymorphism sites of the ethanol dehydrogenase ADH1B and the acetaldehyde dehydrogenase ALDH2 and sequencing primers. According to the method, primer and detection kit for polymorphism detection of the two genes of the ethanol dehydrogenase ADH1B and the acetaldehyde dehydrogenase ALDH2 which are relative to alcohol metabolism, a Sanger sequencing method is adopted, and the primer is used for fast detecting mutation situations of the polymorphism sites of the ethanol dehydrogenase ADH1B and acetaldehyde dehydrogenase ALDH2.

The invention discloses a nucleic acid composition for detecting ALDH2 gene mutation and its application and kit and belongs to the technical field of gene detection. The nucleic acid composition comprises one or more of upstream primers shown in the formulas of SEQ ID NO. 1 and 2, a downstream primer shown in the formula of SEQ ID NO. 3 and a capture probe shown in the formula of SEQ ID NO. 4. The end 5' of the downstream primer or the end 5' of the upstream primer is labeled with an affinant for bonding with a catalytic enzyme. The nucleic acid composition can detect G to A mutation of the site rs671 of the ALDH2 gene and has the characteristics of high sensitivity, high specificity, low cost and convenient operation.

The invention discloses a kit for detecting the sensibility of an individual to glonoine medicines. The kit comprises specific primer pair for detecting SNP polymorphism genotype rs671 on ALDH2 gene, a specific fluorescent probe pair and a routine component used for fluorescent quantitative PCR detection. The sensibility of the individual to the glonoine medicines is analyzed and evaluated by detecting the genotype of SNP locus rs671on the ALDH2 gene.