624 results about "Conserved sequence" patented technology

Method for detecting and identifying unknown bioagents, including bacteria, viruses and the like, by a combination of nucleic acid amplification and molecular weight determination using primers which hybridize to conserved sequence regions of nucleic acids derived from a bioagent and which bracket variable sequence regions that uniquely identify the bioagent. The result is a “base composition signature” (BCS) which is then matched against a database of base composition signatures, by which the bioagent is identified.

The invention relates to a biological detection reagent capable of discriminating three types of ruminant animal varieties simultaneously, a preparation method and an application thereof. The invention selects the specificity of the cow, goat and sheep, and the conservative sequence segment of conservative mitochondrial gene as target, applies Primer Eexpress 3.0 software and Primer Select software in DNAStar, and designs and synthesizes a plurality of groups of primers and probes. After being synthesized and marked, the plurality of pairs of designed primers and the probes are carried out best pairing screening experiment so as to obtain the fittest combination of primers and probes. The reagent contains two pairs of specificity primers, and three Taq Man probes, wherein one pair of theprimer aims at cow source components, the other pair of primer aims at common primers of goats and sheep source component detection; the three probes are respective source components to the cows, thegoats and the sheep; and the amplified target fragment length to the source component detection of the cows, the goats and the sheep is respectively 92bp, 136bp and 136bp. The triple Taq Man fluorescence PCR detection method can simultaneously carry out accurate quantitative detection to the source components of the cows, the goats and the sheep, has simple and convenient operation, intuitive result, strong specificity, high sensitivity and good repeatability and can realize high throughput detection of various source components.

Methods for detecting and identifying unknown bioagents, including bacteria, viruses and the like, by a combination of microRNA containing nucleic acid amplification and molecular weight determination using primers which hybridize to conserved sequence regions of microRNA containing nucleic acids derived from a bioagent and which bracket variable sequence regions that uniquely identify the bioagent. The result is a “base composition signature” (BCS) or molecular mass which is then matched against a database of base composition signatures or molecular masses, by which the species of the bioagent is identified.

Provided are activated collagen scaffold materials as well as their special fused active restoration factors useful for promoting tissue repair, such as bone damage repair or nerve injury repair. The special fused active restoration factors are fusion proteins comprising a collagen-binding domain (CBD) at N- / C-terminus of cytokines, wherein the collagen-binding domain is a polypeptide consisting of 7-27 amino acid residues with a conservative sequence shown in SEQ IN NO:1 at N-terminus.

The invention discloses dsRNA (double-stranded ribonucleic acid) and application to aphid growth inhibition. The dsRNA provided by the invention is dsRNA as expressed by 1) or 2) as follows: 1) dsRNA consisting of ribonucleic acid as shown by a sequence 4 in a sequence table and ribonucleic acid as shown by a reverse complementary sequence of the sequence 4; and 2) dsRNA consisting of ribonucleicacid as shown by a sequence 5 in the sequence table and ribonucleic acid as shown by a reverse complementary sequence of the sequence 5. According to experiments, the obtained dsRNA of a conserved sequence of cytochrome P450cDNA (complementary deoxyribonucleic acid) of English grain aphids and green peach aphids can inhibit growth and development of the English grain aphids and the green peach aphids and can cause a lethal effect by adopting a method of feeding the dsRNA in vitro and using an RNAi (ribonucleic acid interfere) technology to silence the in-vivo cytochrome p450 of the English grain aphids and the green peach aphids.

The present invention is directed to methods of treating flavivirus mediated diseases using siRNAs. The invention is based upon our findings in a mouse model that siRNAs directed against sequences conserved among multiple flaviviruses prevents and treats flavivirus infections. Accordingly, the present invention provides an isolated siRNA comprising a sense RNA and an antisense RNA strand or a single strand. The sense and the antisense RNA strands, or the single RNA strand, form an RNA duplex, and wherein the RNA strand comprises a nucleotide sequence identical to a target sequence of about 15 to about 30 contiguous nucleotides in flavivirus mRNA or mutant or variant thereof.

The invention relates to an EML4-ALK (echinoderm microtubule associated protein like4-anaplastic lymphoma kinase) fusion gene fluorescent quantitative PCR (polymerase chain reaction) assay kit, which is applicable to assay of EML4-ALK fusion gene mutation in lung adenocarcinoma. The kit comprises probes, primers and positive controls, which are specially designed for conserved sequences of 9 fusion variations of the EML4-ALK fusion gene. The kit can be used for quickly and accurately assaying 9 most common EML4-ALK fusion gene variations with high sensitivity, namely 9 fusion variations of 7 variant subtypes, which are subtype 1 (E13; A20), subtype 2 (E20; A20), subtype 3 (E6a / b; A20), subtype 4 (E14; A20), subtype 5 (E2a / b; A20), subtype 6 (E18; A20) and subtype 7 (E14; A20), so that a real-time fluorescent quantitative PCR system for assaying 9 most common EML4-ALK fusion gene variations can be established.

The invention provides a fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) kit for detecting an influenza A virus subtype H7N9. The fluorescent quantitative RT-PCR kit can be used for detection of influenza A viruses and the influenza A virus subtype H7N9. The fluorescent quantitative RT-PCR kit comprises a quantitative RT-PCR reaction solution, an enzyme mixed liquor, a primer and probe mixed liquor, standard substances of influenza A viruses, H7, N9 and RNaseP, positive reference substances of influenza A viruses, H7, N9 and RNaseP), and negative reference substances. Specific primers and probes are designed according to conserved sequences of influenza A viruses, H7 and N9. The RNaseP primers and probes are used as internal references. Through the one-step quadruple real-time fluorescent RT-PCR technology, the influenza A virus and the influenza A virus subtype H7N9 in the sample can be fast and accurately detected. The fluorescent quantitative RT-PCR kit has a reasonable design, very high singularity, sensitivity and repeatability, can be used for laboratory emergency diagnosis and fast screening of an epidemic disease caused by the influenza A virus subtype H7N9, and for an epidemiology study on the influenza A virus and the influenza A virus subtype H7N9 causing fever and respiratory tract syndrome.

The invention discloses a method for detecting bacillus anthracis by combining RPA with a CRISPR technology and a complete set of reagents. The invention provides a complete set of primer group for detecting the bacillus anthracis and / or screening bacillus anthracis virulent strains. The complete set of primer group contains three sets of primers specific to chromosome genes BA_5345 and plasmid virulent genes pagA and capA of the bacillus anthracis virulent strains; each set of primer is composed of an RPA primer and crRNA; the RPA primer is designed according to a target gene conserved sequence; the crRNA comprises an anchoring sequence capable of being combined with cas protein and a spacer sequence matched with an RPA primer amplification product sequence. The complete set of primer group has relatively good sensitivity and specificity for sample detection of clinical simulation samples and soil simulation samples, can effectively distinguish the bacillus anthracis virulent strainsfrom other bacteria, provides a rapid detection method for bacillus anthracis in-vitro diagnosis, and has important significance for soil environment screening of the bacillus anthracis in Chinese epidemic areas.

The invention discloses a method for obtaining an EST-SSR mark, comprising the following steps: (1) obtaining an EST sequence containing simple repeat sequence in genome; (2) in the EST sequence which contains the simple repeat sequence and is obtained in the step (1), classifying the EST sequences with the same simple sequence repeat unit into a same type; (3) performing sequence splicing on the EST sequences of the same type obtained in the step (2) to obtain an overlapping group with variable numbers of simple sequence repeat units, an overlapping group without variable numbers of simple sequence repeat units and an EST sequence without overlapping groups; (4) designing primers according to a side-vane conserved sequence of simple repeat sequence in the overlapping group with available numbers of simple sequence repeat units in the step (3), and detecting the polymorphism of the primers to obtain polymorphic primers, i.e. EST-SSR mark. Compared with the conventional method, the invention increases the development efficiency by 2-4 times, and reduces the work capacity and expenditure, thereby shortening the development time, reducing the development cost and simultaneously reducing the possibility of missing the locus of polymorphism SSR.

The present invention relates to four dodecapeptide sequences capable of binding specifically with surface antigen of tumor metastasis cell, and three dodecapeptides capable of binding specifically with tumor metastasis cell have the following conservative: Pro-Trp-X1-Glu-Pro-X2-Tyr, where X1 and X2 represses any one amino acid each. Owing to the characteristic of specific binding with tumor metastasis cell, these dodecapeptides and their derivatives may be used as the marker for tumor metastasis diagnosis and as the precursor for developing tumor metastasis inhibiting medicines.

The invention belongs to the technical field of biological detection and discloses fluorescent quantitative PCR primers and a kit for detecting novel SADSCoV (swine acute diarrhea syndrome coronavirus). The specific primers and probe are designed according to a nucleocapsid protein gene of a conserved sequence of the SADSCoV, and sequences of the primers and the probe are shown as SEQ ID NO:4-6; the SADSCoV can be amplified specifically by the primers and the probe, for real-time fluorescent quantification, fluorescent data can be acquired by monitoring specific binding condition of the probewith PCR amplified fragments in a PCR process in real time, and the SADSCoV is identified according to Cq value. The SADSCoV can be identified rapidly after PCR amplification with the method, the method is high in accuracy and specificity and good in repeatability and can identify the SADSCoV accurately, rapidly and efficiently, and popularization and application in clinical practice are facilitated.

The invention relates to a CRISPR detection primer set for MTBC (Mycobacterium Tuberculosis Complex) and use thereof, and belongs to the technical field of gene detection. The primer set includes an amplification primer pair and a crRNA; the amplification primer pair is used to amplify a common conserved sequence of M. tuberculosis, M. bovis, M. africanum, M. canettii, and M. microti; the crRNA includes an anchor sequence and a guide sequence, wherein the anchor sequence is bound to a cas protein, and the guide sequence is matched with a targeted RNA fragment in the common conserved sequence.Through the primer set based on CRISPR detection technology, the detection of common conserved sequences of M. tuberculosis, M. bovis, M. africanum, M. canettii, and M. microti by CRISPR detection canquickly perform on-site detection of MTBC, and have the advantages of high specificity, high sensitivity and simplicity.

The invention discloses a PCR (Polymerase Chain Reaction) primer for detecting and identifying porcine circovirus 3 (PCV3) and a detection method and a detection kit. A pair of PCR primers for detecting the PCV3 is designed by performing preference comparison with genomic sequences of the PCV3 according to a conserved sequence on a cap protein of the PCV3. Sequences of the forward and reverse primers PCV3-F / R of the PCR are sequentially shown as SEQ ID NO.1 to SEQ ID NO.2. The invention further provides a PCR detection method and kit for the PCV3. The primer and the detection method disclosed by the invention are capable of rapidly, conveniently and efficiently detecting and identifying the PCV3, the specificity is high, the sensitivity is high, and the detection method is simple and high-efficiency in operation, is convenient for clinical detection and convenient for developing epidemiological investigation and has great application prospects.

The invention belongs to the technical field of diagnostic reagents, and particularly provides a method for amplifying and genotyping nucleic acid genes of HPV (Human Papilloma Virus) and an assay kit for the same. The method comprises the following steps of: carrying out multiple real-time quantitative fluorescence gene amplifications (QPCR) by using a hybrid primer; quantitatively determining high-risk subtype HPVs by using a quantitative probe; and differentiating HPV 16 and 18 subtypes in types by using a typing probe. The hybrid primer is designed by conservative sequences at two sides of a special area of an E1 gene encoded by the HPV; and the genotyping probe is designed according to a type-specific sequence at the center of the area. Gradient dilutions of recombinant plasmids containing 16 and 18 subtype E1-area genes are taken as standard reference. The kit comprises a cervical brush, a sample storage tube / liquid, a primer, a probe, a Taq enzyme & reaction buffer solution, and a standard contrast reference. The system can be used for quantitatively detecting 13 high-risk subtype HPVs (HPV-16, HPV-18, HPV-31, HPV-33, HPV-35, HPV-39, HPV-45, HPV-51, HPV-52, HPV-56, HPV-58, HPV-59, HPV-68) and differentiating two subtypes HPV-16 and HPV-18 in types.

The present invention relates to construction of hepatitis B virus siRNA expression vector and its antiviral therapeutic application. According to the design principle said construction includes the following steps: respectively selecting nucleotide sequence of HBV envelope protein (MS) coding region 69nt-87nt, polymerase(p) coding region 708nt-726nt and core protein (c) coding region 2052 nt-2070nt, designing correspondent siRNA sequence, using BLAST software to analyze homogenously to confirm that it is HBV highly conservative sequence, then chemically synthesizing single-stranded oligonucleotide, external annearing to form double-stranded DNA, connecting it with eukaryotic expression vector pTZU6+1, enzyme excision identification and sequence determination and analysis show that siRNA extression vectors of three target hepatitis B virus (HBV) gene sequences are constructed. The tests show that HBV siRNA has the strong action of inhibiting virus replication and expression.

The invention belongs to the field of biotechnology application and relates to a multiple fluorescence RT-PCR method with internal quality control for simultaneously detecting influenza A virus, influenza B virus and influenza C virus, and a kit. Specific primers and probes are designed for conserved sequences of an M gene, an NS gene and an NP gene of representative strains of the influenza A virus, the influenza B virus and the influenza C virus. The one-step multiple fluorescence RT-PCR method with internal quality control is established. The method is simple and convenient in operation, overcomes tediousness of single detection in single hole of conventional fluorescence RT-PCR detection, simplifies operation processes, and saves the testing cost. The method and the kit provide strong technical support for virus on-site detection, hygienic evaluation of food and water, clinical diagnosis, and the like by utilization of high specificity, high sensitivity, high efficiency and high stability of the method and the kit.

The present invention relates to one pair of PCR primers named as G-dloop, and provides one pair of amplification primers capable of amplifying the high variation zone of rockfish mitochondrial genome in high efficiency and its design method. The amplification primers consist of two single strand oligonucleotides. Through logging-on Genebank to search vertebrate mitochondrion DNA cytochrome b gene and the high conservation area of 16S rRNA gene sequence and homologous comparison, one pair of amplification primers is obtained. Through long PCR amplification, the target segments of 32 varieties of rockfish are obtained and sequenced. The obtained sequences are compared by means of using homologous comparison software Clustal X 1.83 to fine the conservation sequences in the cytochrome b5' ends and the 12S rRNA 3' ends of the mitochondrial genomes of the 32 varieties of rockfish, and the said amplification primers are designed based on the degeneracy principle.

A nerve growth factor with specific combination power to collagen features that a collagen combining structure domain is fused at the amino terminate of its beta subset. It can be used for treating the nerve injury. Its coding gene is also disclosed.

The invention discloses a method for real-time fluorescence isothermal quantitative detection of citrus huanglongbing. The method includes the steps: pretreating a detected sample, and rapidly extracting DNA; according to an HLB16SrDNA conserved sequence, designing two pairs of specific primers including an inner side primer HLB-F3, an inner side primer HLB-B3, an outer primer HLB-FIP and an outer primer HLB-BIP, and preparing the specific primers; carrying out a fluorescence nucleic acid isothermal amplification detection technology reaction; constructing a fluorescence nucleic acid isothermal amplification detection standard curve according to a relationship between plasmid concentrations of a template pMD18-T-HLB with different dilution and corresponding Tt values, wherein the X axis represents log values of starting template concentrations, and the Y axis represents the time taken for amplifying the different-concentration templates to reach thresholds; adopting quantitative detection result judgment, after ending an ESE-Quant TubeScanner reaction, displaying quantitative results by an instrument automatically according to the standard curve, after ending the reaction, instantaneously centrifuging to mix SYBRGreenI on an inner cover of a reaction tube into a reaction liquid, and judging results by adopting color development without opening the cover. The method has the advantages of high sensitivity, high specificity, low pollution, and stable reaction.

The invention provides a primer and a probe for detecting drug resistance genes mecA in methicillin-resistant staphylococcus aureus (MRSA). The invention also provides a method and a detection kit for detecting drug resistance genes mecA in MRSA. According to the invention, the conserved sequence part of drug resistance genes mecA is directly detected without being affected by the mecA gene variation of strains, and a result is a gold standard of MRSA diagnosis. The specific primer and the probe provided by the invention are applicable to currently known strains containing sequence variation.

The invention relates to a rapid screening and detection technology for glyphosate-tolerant transgenic soybean and processed products, in particular to primer groups for detecting the commonly used inserted gene EPSPS of glyphosate-tolerant transgenic plants. Aiming at the EPSPS gene and according to the conserved sequence of the EPSPS gene, the invention designs two groups of specific primers. By adopting the two groups of specific primers and the loop-mediated isothermal nucleic acid amplification technology, the EPSPS gene can be rapidly, sensitively and specifically detected, so the products containing glyphosate-tolerant transgenic plant ingredients are screened. The primer can be provided with other reagents in the form of reagent kits and is used for nucleic acid amplification reaction. The method has the advantages of simple and convenient operation and good repetitiveness.

The invention discloses a triple real-time fluorescent quantitative PCR (polymerase chain reaction) detection method of three simian retroviruses. The method comprises: using conserved sequences from gene gag of simian immunodeficiency virus SIV, gene 5'LTR of Simian T-lymphotropic virus type I to synthesize their DNA fragments, designing and synthesizing three primer pairs and three probes, respectively: SIV: upper primer SI-F, lower primer SI-R and probe SI-P, SRV1: upper primer SR-F, lower primer SR-R and probe SR-P, and STLV1: upper primer ST-F, lower primer ST-R and probe ST-P, and establishing the triple real-time fluorescent quantitative PCR detection method to precisely and quantatively detect three simian retroviruses. The invention also relates to a kit for detection, having the advantages of high detection specificity, high accuracy and good sensitivity.

The invention relates to a multiple RT-PCR method for fast detecting four kinds of pepper viruses, and belongs to the field of virus detection.According to the method, specific primer pairs, namely, CMV-F / R, PMMoV-F / R, BBWV-F / R and TMV-F / R are designed and synthesized aiming at the nucleotide conserved sequences of the four kinds of viruses, multiple RT-PCR system optimization is carried out on the primer concentration, Mg2+ concentration, Taq DNA polymerase concentration, dNTPs concentration, annealing temperature and other aspects influencing multiple RT-PCR amplification, and the detecting method capable of detecting field samples compositely infected by the viruses of CMV, PMMoV, BBWV and TMV in pepper tissue at the same time is built.According to the multiple RT-PCR detecting method, the pepper poisoned condition can be detected fast, accurately, conveniently and economically so that effective prevention measures can be taken as early as possible, and very important significance is achieved on control of diseases in the field, reduction of economic losses, screening of disease resistance and predication of disease epidemiology.

The invention belongs to the field of epizootiology detection, and discloses a goose astrovirus loop-mediated isothermal amplification detection primer group and a kit. According to the method, primers are designed according to goose astrovirus genome conserved sequences: outer primers F3 and B3; inner primers FIP and BIP; a loop primer LB. The specific sequences are shown in SEQ ID NO.1-5 separately. According to the designed primers, a loop-mediated isothermal amplification method for detecting goose astroviruses is established. According to the detection method, no cross reaction with common goose common goose infectious disease pathogens occurs. The detection method is high in specificity, good in accuracy, high in sensitivity, free of expensive instruments and equipment and convenient and facilitates public use.

The invention belongs to the field of gene engineering and genetic modification and relates to construction of a CRISPR-Cas9 system of a tomato PSY 1 gene and an application thereof. The construction includes the following steps: (1) according to a cDNA conserved sequence of the tomato PSY 1 gene, designing a DNA sequence of sgRNA, which is represented as the SEQ ID No.1; (2) designing a primer combination used for amplifying a tomato PSY 1 gene target zone in the sgRNA; and (3) performing sgRNA amplification with the primer and purifying an amplified product to obtain a CRISPR-Cas9 Level-1 carrier and a CRISPR-Cas9 Level-2 carrier, which contain the sgRNA of the specific targeted tomato PSY-1 gene. The construction is beneficial to researching on effect that the gene participates in fruit maturity of tomatoes, so that the molecular mechanism of the fruit maturity of tomatoes can be known furthermore. The system can be used for producing PSY 1 gene deleted transgenic tomatoes, and has important effect of culturing a new tomato variety which is tolerant to storage and transportation in future.

The invention provides a gene conserved sequence, a primer probe, a kit and application for detecting neocoronavirus, and belongs to a molecular biological nucleic acid detection technology. In orderto solve the technical problems of better specific detection of the neocoronavirus, sensitivity during detection and the like, the invention provides the gene conserved sequence, the primer probe, thekit and the application for detecting an ORF1ab / N gene of the neocoronavirus, and belongs to the molecular biological nucleic acid detection technology. The invention aims to improve the specificity,the sensitivity and the detection efficiency of neocoronavirus detection, so that the whole detection process is simpler, more convenient, quicker and more efficient.

The invention provides a conserved sequence of locust's ATP synthase beta subunit cDNA, and constructs the RNA, DNA and other constructs thereof. The RNAi technology is utilized to silence the ATP synthase beta subunit in locusts, so that expression of in vivo ATP synthase beta subunit can be significantly inhibited, thus resulting in the lethal effect of locusts. The Locusta migratoria ATP synthase beta subunit gene and its dsRNA can be applied to the RNAi technology for control of Locusta migratoria pests.

The invention relates to an alkalic xylanase gene and an engineering bacterium containing the same. The alkalic xylanase gene has the overall length of 687 bp, is called XynG1-3, encodes 228 amino acids and predicts that the molecular weight of the amino acids is 25444Da, wherein the first 27 amino acids are signal peptides of the alkalic xylanase gene, and 38th-221st amino acids are conserved sequences of a glycoside hydrolase GH11 family. Compared with a reported xylanase gene sequence, individual nucleotides of the obtained and expressed alkalic xylanase gene are different; the optimal operative temperature and the optimal pH of the recombinase are respectively 55 DEG C and 8.0, relative enzyme activity is more than 50 percent after preserving heat at the pH of 9.0 and 60 DEG C for 1 hour, and the relative enzyme activity is more than 70 percent after preserving the heat under the conditions of the pH of 6-9 and the temperature of 55 DEG C for 1 hour.