621 results about "Conserved sequence" patented technology

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.

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 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 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 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 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 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 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.