45 results about "Intergenic region" patented technology

The invention provides a recombinant vector for eliminating the activity of a kanamycin drug resistance gene, and aims at eliminating drug resistance germs in organisms and solving the problem of kanamycin drug resistance of the germs. The recombinant vector for eliminating the activity of the kanamycin drug resistance gene is characterized by comprising a pCas9 vector subjected to chloramphenicol resistance elimination and a gRNA nucleotide sequence KR58 or KR208 aiming at a kanamycin resistance gene kan; the concrete nucleotide sequence of the KR58 is GCCGCGAT TAAATTCCAACA, and the concrete nucleotide sequence of the KR208 is CAATGATG TTACAGATGAGA. A building method of the recombinant vector mainly comprises the steps of carrying intergenic region nucleic acids by a novel gene editing tool CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) / Cas9 system; removing a chloramphenicol resistance gene on the recombinant vector; transforming the gene into vaccine vector bacteria such as attenuated salmonella typhimurium; performing co-culture on the recombinant bacteria and the kanamycin drug resistance gene so that the recombinant vector in the recombinant bacterium cell enters the kanamycin drug resistance bacteria in an engaging mode. The activity of the kanamycin resistance gene kan is effectively inhibited, so that the original drug resistance bacterium cannot grow on a kanamycin culture medium.

The invention relates to a method for identifying honeysuckle and lonicera confuse and an application of the method. The method disclosed by the invention comprises the step of performing PCR (polymerase chain reaction) amplification on ITS2 nucleotide sequence, and particularly comprises the steps of 1) extracting the DNA (deoxyribonucleic acid) of a sample; 2) performing PCR amplification on the segments of the ITS2 sequence containing ribosome DNA; 3) splicing the amplification products to obtain a complete ITS2 intergenic region; and 4) establishing the NJ (neighbour-joining) tree of the ITS2 sequence of the sample. The method disclosed by the invention can be used for effectively solving the problems of variety identification, variety improvement and breeding of honeysuckle medicinal materials, development and utilization of germplasm resources, and the like; and the method disclosed by the invention is wide in applicability, simple to operate, easy to grasp, high in accuracy, and capable of successfully realizing rapid and accurate identification on honeysuckle medicinal materials or powder and lonicera confuse medicinal materials or powder.

The present invention relates to novel insertion sites useful for the integration of exogenous sequences into the Modified Vaccinia Ankara (MVA) virus genome. The present invention further provides plasmid vectors to insert exogenous DNA into the genome of MVA. Furthermore, the present invention provides recombinant MVA comprising an exogenous DNA sequence inserted into said new insertion site as medicine or vaccine.

The invention provides a tubercle bacillus drug tolerance detection reagent kit and method. The tubercle bacillus drug tolerance detection reagent kit comprises a tubercle bacillus drug tolerance detection reagent, wherein the tubercle bacillus drug tolerance detection reagent comprises a sequencing primer in accordance with a tubercle bacillus drug tolerance gene; the tubercle bacillus drug tolerance gene comprises one or more genes of rpoB, katG, inhA-promoter, inhA-structural, furA, embB, ubiA, pncA, rpsA, gyrA, gyrB, eis, rpsL, rrs, tlyA, rplC and rrl; and further, the reagent kit also contains a tubercle bacillus nucleic acid detection reagent, and the tubercle bacillus nucleic acid detection reagent comprises a primer pair 1 in accordance with IS6110, a primer pair 2 in accordance with the IS6110 and a probe primer in accordance with the IS6110. Through the adoption of the tubercle bacillus drug tolerance detection reagent kit disclosed by the invention, tubercle bacillus nucleicacid in samples can be quickly detected, and positive samples can be further subjected to drug tolerance detection; and the tubercle bacillus drug tolerance detection reagent kit has good sensitivity, good specificity and good accuracy, and can perform mutation detection on 48 sites of 17 drug tolerance genes of common antituberculosis drugs and fragment deficiency detection of an intergenic region, so that the tuberculosis medication can be more accurately and comprehensively guided.

The present invention relates to recombinant modified vaccinia Ankara (MVA) virus containing restructured sites useful for the integration of heterologous nucleic acid sequences into an intergenic region (IGR) of the virus genome, where the IGR is located between two adjacent, essential open reading frames (ORFs) of the vaccinia virus genome, wherein the adjacent essential ORFs are non-adjacent in a parental MVA virus used to construct the recombinant MVA virus, and to related nucleic acid constructs useful for inserting heterologous DNA into the genome of a vaccinia virus, and further to the use of the disclosed viruses as a medicine or vaccine.

The invention provides an improved PCR detection method of the dwarfing bacteria of sugarcane persistent roots. A pair of specific primers Lxx1 / Lxx2 are designed according to an RSD pathogenic bacterium Lxx 16S-23SrDNA intergenic region nucleotide sequence, and a PCR multiplication method is adopted to detect the dwarfing bacteria of sugarcane persistent roots after the improved method is used for extracting the total DNA of sugarcane juice; the method overcomes the defects of the prior detection technique; and the invention provides the PCR detection method of the dwarfing bacteria of the sugarcane persistent roots, which has the advantages of high speed, stability, accuracy, high sensitivity and strong specificity.

The invention discloses a new mutation site related to isoniazide resistance of mycobacterium tuberculosis and an application thereof. The new mutation site is positioned at a 1673425th site of a standard strain H37Rv genome, that is a 126th base in an intergenic region of Rv1482c to Rv1483*, and the generated point mutation comprises that a base C is mutated into a base T or a base A. The found new mutation site can be used as a detection target applied in detection of the isoniazide resistance of mycobacterium tuberculosis, improves the molecular detection level of the isoniazide resistance of mycobacterium tuberculosis, shortens the diagnosis time of patients, and saves the treating time and cost for the patients. The mycobacterium tuberculosis drug-resistant mutation site is explained to possibly occur in a non-coding region, the base mutation of the intergenic region is prompted to be possibly related to the drug resistance, and a drug-resistant biomarker also exists. The invention provides a new idea for finding out a new and more effective mycobacterium tuberculosis drug-resistant molecular marker.

The invention discloses a new mutation site related to streptomycin resistance of mycobacterium tuberculosis and an application thereof. The new mutation site is positioned at a 1338631st site of a standard strain H37Rv genome, that is a 47th base in an intergenic region of Rv1194c to Rv1195, and the generated point mutation comprises that a base C is mutated into a base A. The found new mutation site can be used as a detection target applied in detection of the streptomycin resistance of mycobacterium tuberculosis, improves the molecular detection level of the streptomycin resistance of mycobacterium tuberculosis, shortens the diagnosis time of patients, and saves the treating time and cost for the patients. The mycobacterium tuberculosis drug-resistant mutation site is explained to possibly occur in a non-coding region, the base mutation of the intergenic region is prompted to be possibly related to the drug resistance, and a drug-resistant biomarker also exists. The invention provides a new idea for finding out a new and more effective mycobacterium tuberculosis drug-resistant molecular marker.

The present invention relates to detection of pathogenic mycobacteria in clinical specimens such as sputum, cerebrospinal fluid, gastric lavage and tissue biopsies etc., wherein the novel stretch of DNA that lies in the intergenic region between methyl mycolic acid synthase genes mmaA1 and mmaA2 and the flanking region in mmaA1 and mmaA2 genes and is the invention uses a pair of designed oligonucleotide primers that specifically amplifies the target DNA from the clinical specimens

The invention belongs to the field of biochemistry, and relates to an MEV (mevalonic acid) pathway tuned by TIGR (tunable intergenic regions) to produce isopentene compounds, especially zeaxanthin. In particular, the exogenous MEV pathway is introduced into the engineered escherichia coli (ZEAX) for producing zeaxanthin, especially an upstream pathway and a downstream pathway of MEV tuned by the TIGR are introduced to coordinate the production of intermediate products and reduce the toxicity to cells due to the accumulation of intermediate products, and finally the yield of zeaxanthin is increased.

The invention relates to a heat-resistant serum 4 type fowl adenovirus genetic engineering vaccine candidate strain and a construction method thereof. The candidate strain is a thermal-stability newcastle disease virus (Newcastle Disease Virus) rAHR09-4F2 expressing serum 4 type fowl adenovirus spike protein 2, and the preservation number is CCTCC NO: V201932. The candidate strain is obtained through using a heat-resistant newcastle disease attenuated virus strain rAHR09 as a carrier, inserting a serum 4 type fowl adenovirus (FAdV4) spike protein (fiber2) gene between a P intergenic region andan M intergenic region, and adopting a backward heredity technique. The result of evaluating the biological characteristics and the immunoprotection efficacy of the candidate strain displays that therecombinant virus has favorable heat stability and immunogenicity and provides favorable immunoprotection on FAdV4 infection.

The present invention relates, in one aspect, to a method for determining the severity of a disease attributed to at least one genetic mutation in one or more of the genes encoding haemoglobin polypeptide chains, comprising the steps of: (a) providing a sample from said subject; and (b) determining the presence of one or more diagnostic markers:(i) within a 127 kb segment on chromosome 2p15;(ii) within MYB and / or HBSIL and / or the intergenic region between MYB and HBSIL located on the 6q23 QTL interval; and / or(iii) within one of the chromosomal loci given in Table 14; wherein the presence of said marker(s) in said sample is indicative that the severity of said disease in said subject will be or is less severe in said subject in comparison to a subject that does not possess said marker(s).

Intergenic non-coding RNA molecules that regulate the expression of HWP1 and ALS3 of Candida are provided as are methods of using the non-coding RNA molecules and complementary molecules thereof in modulating HWP1 or ALS3 expression; adherence, yeast-to-hyphal transition, or biofilm development of Candida; and preventing or treating candidiasis.

The invention relates to a method for identifying honeysuckle and lonicera confuse and an application of the method. The method disclosed by the invention comprises the step of performing PCR (polymerase chain reaction) amplification on ITS2 nucleotide sequence, and particularly comprises the steps of 1) extracting the DNA (deoxyribonucleic acid) of a sample; 2) performing PCR amplification on the segments of the ITS2 sequence containing ribosome DNA; 3) splicing the amplification products to obtain a complete ITS2 intergenic region; and 4) establishing the NJ (neighbour-joining) tree of the ITS2 sequence of the sample. The method disclosed by the invention can be used for effectively solving the problems of variety identification, variety improvement and breeding of honeysuckle medicinal materials, development and utilization of germplasm resources, and the like; and the method disclosed by the invention is wide in applicability, simple to operate, easy to grasp, high in accuracy, and capable of successfully realizing rapid and accurate identification on honeysuckle medicinal materials or powder and lonicera confuse medicinal materials or powder.

The invention provides a gene chip and a kit for detecting common pathogenic bacteria in food. The gene chip comprises a solid phase carrier and an oligonucleotide probe fixed on the solid phase carrier, wherein the oligonucleotide probe comprises DNA sequences which are selected from an nuc gene of staphylococcus aureus, an speB gene of streptococcus pyogenes, an invA gene of salmonella, an ipaHgene of Shigela, a zpx gene of Enterobacter sakazakii, a 16S-23S intergenic region of Klebsiella pneumoniae, a toxR gene of Vibrio parahaemolyticus and 16SDNA of the seven bacteria. The kit comprisesthe gene chip. By utilizing the gene chip and the kit to detect the seven common pathogenic bacteria in food, the invention has the advantages of simple and convenient operation, high accuracy and good repeatability.

The invention relates to a novel isothermal loop-mediated vibrio parahaemolyticus nucleic acid marker detection reagent. According to the reagent, an antigen is marked for a front inner primer in a primer group used in isothermal loop-mediated amplification, meanwhile another antigen is marked for a rear inner primer, the operation can be performed when a vibrio parahaemolyticus specific 16S-23S rRNA intergenic region sequence is amplified, subsequently a piece of colloidal gold test paper in match is adopted to detect an amplification product, and then vibrio parahaemolyticus can be detected. As the primer for the 16S-23S rRNA intergenic region sequence is designed for RNA secondary structure prediction, the situation that the detection sensitivity and the specificity can be influenced by mutation caused if other protein genes are used or the rRNA gene is blindly used can be avoided as much as possible, and the detection reagent provided by the invention is good in specificity and sensitivity.

The invention provides an SNP marker of a medicinal plant murraya paniculata. In the SNP marker, the nucleotide on the 75th site from the 5' terminal in the sequence shown by SEQ ID No.1 is T, the nucleotide on the 109th site is T and / or the nucleotide on the 173rd site is A; the sequence shown by SEQ ID No.1 is the ITS2 intergenic region sequence of murraya paniculata. By adopting the SNP marker, the medicinal murraya paniculata can be quickly and accurately identified; the SNP marker has wide applicability and can guarantee the stability and reliability of the identification result from the source; and compared with traditional morphological identification method, the SNP marker of the medicinal plant murraya paniculata can realize quick and accurate identification of the medicinal murraya paniculata.

The invention discloses a quick identification method for a paniculate swallowwort root medicinal material and counterfeit species thereof, namely rhizoma cynanchi stauntonii and blackend swallowwortroots. The quick identification method comprises the following steps: 1) extracting DNA of a to-be-detected sample; 2) taking the DNA of the to-be-detected sample as a template, and performing PCR amplification on a fragment of an ITS2 sequence containing ribosomal DNA; 3) splicing amplification products to obtain a complete ITS2 intergenic region; 4) analyzing interspecific variation sites of paniculate swallowwort root and counterfeit species thereof, namely rhizoma cynanchi stauntonii and blackend swallowwort roots, identifying as paniculate swallowwort roots if the 50-th site is a basic group C, and identifying as counterfeit species of paniculate swallowwort roots if the 50-th site is basic group deletion. The quick identification method is strong in applicability, is simple to operate, can realize quick and accurate identification for the paniculate swallowwort root medicinal material and counterfeit species thereof, and can identify a great number of samples within a short time.

This invention relates to a method for stably expressing a transgene integrated into the genome of an animal cell or of an animal over a long period. Specifically, this invention provides: an approximately 2.5 kb XhoI-BamHI fragment (or XB fragment) derived from the Evx2-Hoxd13 intergenic region of the animal genome, or a homologue thereof; a DNA containing a foreign DNA wherein the DNA has been inserted between the two essentially identical XB fragments or homologues thereof; a vector, animal cell, or nonhuman mammalian animal containing said DNA; and use of the vector, animal cell, or nonhuman mammalian animal for production of a substance or therapy.

The invention discloses a molecular marker primer and a method which are used for identification of prunus pedunculata, prunus mongolica, and prunus triloba. The molecular marker primer comprises a forward primer and a reverse primer; the sequence of the forward primer tKF is ACTCGAACCCGGAACTAG, and the sequence of the reverse primer tKR is CGACCGATTTCTGCGTAT. The identification method comprises following steps: 1, leaf sample collecting; 2, leaf sample total genome DNA extraction; 3, amplification of target genes in leaf sample total genome DNA obtained in step 2 using the molecular marker primer used for identification of prunus pedunculata, prunus mongolica, and prunus triloba, and obtaining of amplification products; 4, purifying and sequencing of the amplification products obtained instep 3; and 5, comparison analysis on sequencing results obtained in step 4. According to the method, trnK-UUU intergenic region based base difference can be taken as the molecular marker primer usedfor identification of prunus pedunculata, prunus mongolica, and prunus triloba germplasm distinguishing, the identification method is simple, and operation is convenient.

The invention belongs to the technical field of algae molecular markers, and particularly relates to a mitochondria molecular marker primer of enteromorpha population and application thereof. The mitochondria molecular marker primer is applied to identify a high-resolution mitochondria molecular marker (the sequence of a rps2-trnL intergenic region) of the enteromorpha population. The method can detect an algae sample and a microcosmic propagule sample, and is more reliable and practical compared with a conventional method, and accordingly the application of the mitochondria molecular marker primer of the enteromorpha population has significant meaning on the aspects of green tide disaster research, enteromorpha germplasm identification, phylogeny, systematic geography research and protection and utilization of enteromorpha resources.

The invention discloses a molecular marker closely linked with a tomato neck and root rot resistant gene Frl, and application of the molecular marker. The invention provides application of a substance for detecting whether a 165bp fragment is contained in a positioning interval of an Frl gene in the genome of a tomato variety or not in identification or auxiliary identification of tomato neck and root rot resistance of tomatoes. The invention discovers that 165bp large fragment insertion related to the tomato neck and root rot resistance is in the presence in an intergenic region in the positioning interval (an interval of 4.2-5.1 Mb of a tomato No.9 chromosome) of the Frl gene in the genome of the tomato variety, a PCR-based molecular marker is designed according to the fragment, whether a tomato material to be detected contains the Frl gene or not and whether the tomato material is homozygous or not can be detected, a breeding period can be shortened, and the accuracy of breeding selection is improved.

A method for the high level production of active, properly processed recombinant protein in trans-splicing organisms is disclosed. The method involves the integration of the gene encoding the recombinant protein of interest into a chromosomal locus where it is transcribed under the direction of the rRNA promoter. The gene is also operably linked to intergenic regions allowing the protein to be translated in these organisms. The recombinant organisms expressing a therapeutic protein can also be used to treat a disease or undesirable condition which is characterized by a deficiency in that protein.