10037 results about "Nucleotide sequencing" patented technology

Localized detection of RNA in a tissue sample that includes cells is accomplished on an array. The array include a number of features on a substrate. Each feature includes a different capture probe immobilized such that the capture probe has a free 3′ end. Each feature occupies a distinct position on the array and has an area of less than about 1 mm2. Each capture probe is a nucleic acid molecule, which includes a positional domain including a nucleotide sequence unique to a particular feature, and a capture domain including a nucleotide sequence complementary to the RNA to be detected. The capture domain can be at a position 3′ of the positional domain.

Localized detection of RNA in a tissue sample that includes cells is accomplished on an array. The array include a number of features on a substrate. Each feature includes a different capture probe immobilized such that the capture probe has a free 3′ end. Each feature occupies a distinct position on the array and has an area of less than about 1 mm2. Each capture probe is a nucleic acid molecule, which includes a positional domain including a nucleotide sequence unique to a particular feature, and a capture domain including a nucleotide sequence complementary to the RNA to be detected. The capture domain can be at a position 3′ of the positional domain.

The invention belongs to the technical field of genetic breeding, and particularly relates to a molecular marker influencing sheep weight traits and application thereof, and the molecular marker comprises a site I or a site II. The nucleotide sequence of the molecular marker containing the site I is as shown in SEQ ID NO.1, and mutation from T to C occurs at the 101bp position in the SEQ ID NO.1; the nucleotide sequence of the molecular marker containing the site 2 is as shown in SEQ ID NO.2, and mutation from A to G occurs at the 101bp position in the SEQ ID NO.2. According to the molecular markers influencing the sheep weight traits, the marker I and the marker II can be used for identifying the sheep weight traits, and the offspring weight can be increased by selecting TT or AA genotype individuals as male parents or female parents.

The invention belongs to the technical field of cosmetics, and discloses anti-aging recombinant collagen as well as a preparation method and application thereof. The amino acid sequence of the anti-aging recombinant collagen is as shown in SEQ ID NO.1, and the nucleotide sequence of the anti-aging recombinant collagen is as shown in SEQ ID NO.2. The recombinant collagen CaI1 is constructed and expressed by recombining the effective sequences of the I-type collagen, the III-type collagen and the XVII-type collagen, and the recombinant collagen CaI1 is free of cytotoxicity, free of hidden danger of endotoxin and good in safety. The skin care product added with the recombinant collagen CaI1 can promote skin collagen absorption, inhibit skin aging and improve skin elasticity, and has a good application prospect.

The invention discloses an AsMYB19 gene and application thereof, and relates to the technical field of biology, the nucleotide sequence of the AsMYB19 gene is shown as SEQ ID NO.1, the AsMYB19 gene can regulate and control the tiller number and the yield of oat, gene resources and strategies are provided for oat breeding and improvement of oat germplasm resources, the breeding speed is increased, and the breeding efficiency is improved.

The invention discloses an AsSN1 gene and application, and relates to the technical field of biology. The nucleotide sequence of the AsSN1 gene is as shown in SEQ ID NO.1, and the amino acid sequence of the AsSN1 protein is as shown in SEQ ID NO.2. The AsSN1 gene and the protein provided by the invention can regulate and control the spikelet number of the oats, further regulate and control the yield of the oats, provide important support for oat breeding and germplasm resource improvement, and have important significance for meeting the development requirements of animal husbandry.

The present invention provides a tobacco fragrance gene, its targeted sgRNA and applications, belonging to the technical field of tobacco breeding. A tobacco fragrance gene includes the NtBADH2a and / or NtBADH2b gene; the nucleotide sequence of the NtBADH2a gene is as shown in SEQ ID NO:1; the nucleotide sequence of the NtBADH2b gene is as shown in SEQ ID NO:2. The application of the tobacco fragrance gene as a target in improving the fragrance of tobacco. In view of the fact that the tobacco fragrance gene affects the content of tobacco fragrance substances through negative regulation, the present invention also provides the application of an sgRNA or gene editing system targeting the tobacco fragrance gene in constructing a tobacco line rich in fragrance substances. It can be seen that the tobacco fragrance gene provided by the present invention provides an effective target for tobacco breeding.

The present invention discloses a mutant strain of Streptomyces parvum and its application in producing glutathione, belonging to the field of genetic engineering. The present invention successfully constructs a mutant strain of Streptomyces parvum by overexpressing the glnA gene in Streptomyces parvum; wherein the nucleotide sequence of the glnA gene is shown in SEQ ID NO: 1. Experiments have shown that compared with the wild-type Streptomyces parvum, the mutant strain of Streptomyces parvum can significantly increase the content of glutathione and glucose in the fermentation product, and can also improve the utilization efficiency of ammonia nitrogen during the fermentation process. The present invention provides new strain resources and methods for mass production of bacterial-derived glutathione, and provides new agricultural antibiotic resources and strategies for preventing and controlling a variety of plant diseases.

The invention discloses a primer combination for detecting monkey pox viruses and identifying monkey pox viruses Ia, Ib and II, and belongs to the technical field of molecular biology, the nucleotide sequences of specific primers and probes are shown as SEQ ID NO.1-SEQ ID NO.12, and the primer combination aims at solving the problem that in the prior art, new evolutionary branch Ib types of monkey pox viruses cannot be accurately typed, and the detection sensitivity is high. According to the present invention, the defects that the monkey pox virus and the orthopox virus have the cross reaction are overcome, the genome difference of different evolutionary branches is analyzed through the bioinformatics system, the primer probe composition capable of simultaneously detecting the monkey pox virus and carrying out accurate typing is analyzed and screened, and the primer combination can detect 200 copies / mL of the monkey pox virus with high specificity and high sensitivity. Meanwhile, detection of the monkey pox virus and identification of the types Ia, Ib and II of the monkey pox virus are realized in a reaction system, so that different monkey pox branches can be distinguished, and treatment and prevention can be performed in a more targeted manner.

The invention discloses a hemoglobin-resistant Taq DNA polymerase mutant and a construction method thereof, and relates to the field of biology, and the hemoglobin-resistant Taq DNA polymerase mutant is characterized in that a nucleotide sequence for coding the Taq DNA polymerase is shown as SEQ ID NO.1, and the mutant comprises at least one mutation or all mutations selected from S623D and E721A sites. On the basis of natural Taq DNA polymerase, the molecular structure of the Taq DNA polymerase is modified through rational design and combination with a site-specific mutagenesis biotechnology, so that the Taq DNA polymerase mutant is more suitable for PCR amplification of samples containing hemoglobin and the like than a wild type, and the result judgment accuracy of a sample containing a hemoglobin inhibitor is more facilitated.

The invention provides application of an ECT2 gene in regulating and controlling development of secondary xylem of a poplar. Transgenic poplar with overexpressed ECT2 genes is obtained through genetic transformation, it is found that overexpressed ECT2 genes can increase the width of secondary xylem of the poplar, and the nucleotide sequence of the ECT2 genes is shown as SEQ ID NO: 1. The ECT2 gene has a regulation effect on the development of the secondary xylem of the poplar.

The invention relates to the technical field of molecular marker development and molecular marker assisted breeding, and particularly discloses a KASP molecular marker closely linked with a wheat drought resistance related gene TaRPL36-8, a primer and application, the nucleotide sequence of the KASP molecular marker is shown as SEQ ID NO.1, and T / C polymorphism exists at the 201bp position of the sequence. By detecting the KASP molecular marker, the drought-resistant variety of wheat is identified, and technical support is provided for wheat breeding.

The invention discloses a PgEGY3 gene for improving cold resistance of pennisetum alopecuroides and increasing fresh weight of leaves and application of the PgEGY3 gene, and relates to the technical field of genetic engineering, and the nucleotide sequence of the PgEGY3 gene of the pennisetum alopecuroides is shown as SEQ ID NO.1. The invention also discloses a protein coded by the Pennisetum alopecuroides PgEGY3 gene. The invention also discloses a recombinant expression vector and a recombinant host cell comprising the Pennisetum alopecuroides PgEGY3 gene. The invention also discloses application of the PgEGY3 gene, a recombinant expression vector and a recombinant host cell in improving cold resistance and biomass of pennisetum alopecuroides. It is verified that the Pennisetum alopecuroides PgEGY3 gene can improve the cold resistance of plants, and the growth state of the plants under cold stress is remarkably improved through overexpression of the PgEGY3 gene.

The embodiment of the invention discloses a KASP molecular marker, a primer pair and a kit for identifying the dwarf character of muskmelon and application of the KASP molecular marker, relates to the technical field of molecular markers, and aims to promote gene positioning and molecular-assisted breeding processes by utilizing molecular marker-assisted selective breeding on the dwarf character of muskmelon. The method for identifying the dwarf character of the muskmelon comprises the following steps: extracting genome DNA of a target muskmelon, taking the genome DNA of the target muskmelon as a template for PCR amplification, and performing fluorescence detection based on a primer of a KASP marker K1069930 to obtain a gene type of the target muskmelon, on the basis of the detected gene type, the character of the target muskmelon is judged; the KASP marker K1069930 is designed according to an SNP (Single Nucleotide Polymorphism) site at Chr07: 1069930, and the nucleotide sequence of the SNP site is shown as SEQ ID NO. 1. The invention also discloses a kit for detecting the KASP marker K1069930 and a kit for detecting the KASP marker K1069930. The identification method provided by the invention can be more efficiently applied to breeding work of dwarf muskmelon germplasm.

The invention discloses a pennisetum purpureum gene PpbHLH148L and application thereof in improving the cold resistance of plants, and relates to the technical field of gene engineering. The nucleotide sequence of the gene is as shown in SEQ ID NO.5. After the gene is subjected to subcellular localization and expression condition analysis after cold stress, the gene is related to low-temperature tolerance. An overexpression vector of the gene is constructed, Agrobacterium tumefaciens is transferred, then Arabidopsis thaliana infection is carried out, cold tolerance determination is carried out on overexpressed Arabidopsis thaliana, phenotypic difference and physiological index difference between an overexpressed strain after cold treatment and a wild type are observed, and it is found that the gene can significantly improve the low-temperature stress tolerance of Arabidopsis thaliana. And in the low-temperature treatment of the pennisetum alopecuroides transgenic callus, the transgenic callus also shows higher tolerance than the wild callus at low temperature. The pennisetum purpureum gene PpbHLH148L can be used for low-temperature tolerance modification of plants, and has an application prospect in creation of low-temperature-resistant plant materials.

The invention discloses a double-dominant SCAR marker for identifying allium fistulosum and allium fistulosum polluted by onion pollen and application of the double-dominant SCAR marker. The specific fragment length of the SCAR marker of an abnormal plant polluted by onion pollen is 1481 bp and 851 bp, the nucleotide sequences of the SCAR marker are shown as SEQ ID NO.1 and SEQ ID NO.2 respectively, the specific fragment length of the SCAR marker of the allium fistulosum is 840 bp, and the nucleotide sequence of the SCAR marker is shown as SEQ ID NO.3; the invention also discloses a specific primer of the SCAR marker. When the marker determined by the method is used for verifying different varieties of green Chinese onions and abnormal plants of the green Chinese onions polluted by the onion pollen, the green Chinese onions and the abnormal plants of the green Chinese onions polluted by the onion pollen can be quickly and stably identified, and compared with a traditional seed purity identification method, the method is not influenced by natural environment and human factors, can be used for batch experiments, improves the identification efficiency, and has a wide application prospect. The invention provides a rapid and feasible method for solving the problem that allium fistulosum varieties are mixed due to the fact that allium fistulosum pollen is polluted by the allium fistulosum pollen cannot be identified in production.

The invention provides a fluorescent protein selection marker staygold-ygjHr gene and application thereof, and belongs to the technical field of biological engineering. The preparation method comprises the following steps: replacing codons of L-valine in nucleotide sequences of a fluorescent protein gene Staygold and a gene ygjH by using a rare codon GTC to respectively obtain a fluorescent protein gene Staygoldr after codon replacement and a gene ygjHr after codon replacement, and connecting the two segments of genes after codon replacement by using a flexible protein peptide to obtain the fluorescent protein selection marker Staygold-ygjHr gene. Experiments prove that the gene can obviously improve the screening efficiency of L-valine high-yield strains.

The invention discloses a target segment for preventing and treating tomato brown fruit wrinkling virus in tobacco, double-stranded RNA (Ribonucleic Acid) and application of the target segment and the double-stranded RNA. Wherein the nucleotide sequence of the target segment is as shown in SEQ ID NO.3, or a sequence which is as shown in SEQ ID NO.3, has at least 90% of homology and has coded amino acid as shown in SEQ ID NO.4 is substituted. The invention also discloses a double-stranded RNA of a targeted interference target section, and the double-stranded RNA can significantly inhibit the proliferation of the tomato brown crinkled fruit virus in tobacco by injecting or smearing tobacco leaves, so that the double-stranded RNA can be effectively applied to prevention and control of tobacco diseases caused by the tomato brown crinkled fruit virus. The method for preventing and controlling the tomato brown crinkled fruit virus in the tobacco by using the double-stranded RNA is safe, harmless and efficient, and provides a new path for preventing and controlling plant virus diseases by using RNAi (Ribonucleic Acid Interference).

The invention provides a primer group for targeted simultaneous detection of multiple bacterial pathogens based on multiple amplicons and application of the primer group, and belongs to the technical field of biological detection of pathogenic bacteria. The primer group for targeted simultaneous detection of multiple bacterial pathogens based on multiple amplicons provided by the invention consists of 196 pairs of primers, and the nucleotide sequences of the primers are shown as SEQ ID NO: 1-392. By using the primer group provided by the invention, multiple pathogens can be rapidly and efficiently detected at the same time, and the primer group has the technical advantages of high targeting, strong specificity and the like.

The invention provides a Ustilaginoidea virens gene UvASE2 for regulating and controlling rice resistance and application of the Ustilaginoidea virens gene UvASE2, and relates to the technical field of plant genetic engineering. The nucleotide sequence of the Ustilaginoidea virens gene UvASE2 is as shown in SEQ ID NO: 1, and the amino acid sequence of the encoding protein of the Ustilaginoidea virens gene UvASE2 is as shown in SEQ ID NO: 2. By constructing a transgenic rice plant for heterologous expression of UvASE2, it is found that the transgenic rice can significantly improve the resistance to false smut and bacterial blight and significantly induce the outbreak of active oxygen. The heterologous expression UvASE2 gene of Ustilaginoidea virens has the function of positively regulating and controlling the disease resistance of rice, and the UvASE2 gene can be used for improving the disease resistance of rice and has important significance on creation of disease-resistant germplasm of rice.

A resistance gene TaCAT to FCR of wheat and an application thereof are provided, which relates to the field of biological genetic engineering technologies. The nucleotide sequence of a genome where the TaCAT gene is located, the nucleotide sequence of a cDNA, the nucleotide sequence of a CDS and a coded protein sequence are respectively as shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4. An InDel marker is developed for identifying different allelotypes of the TaCAT gene. An application of the TaCAT gene for preparation of transgenic plant cells and plant breeding is provided. The molecular genetic basis of the resistance to FCR is disclosed, an important role is played in improving the resistance to FCR by utilizing a genetic engineering technology, and a new way is provided for breeding a new wheat variety with disease-resistance and safety.

The invention provides a production strain of an antibiotic-free plasmid, the production strain is a gene editing strain of a PIR strain and is named as PIR1-WN:: 0636 or PIR1-PR: 0636, the production strain contains a nucleotide sequence for coding toxin protein and the antibiotic-free plasmid, and the antibiotic-free plasmid contains a nucleotide sequence for coding antitoxin protein; and preferably, the replicon DNA element of the nonreactive plasmid is R6K-gamma. The toxin protein gene of the production strain disclosed by the invention can be stably passaged, has lethality after being induced and can be used for plasmid screening; according to the invention, the positive rate of transforming the nonreactive plasmid into the PIR1-WN:: 0636 strain is more than 80%, and stable production of the plasmid with a high superhelix ratio can be realized.

The specification provides methods for introducing a desired genetic change in a nucleotide sequence using a double-strand break (DSB)-inducing genome editing system, the method comprising: identifying one or more available cut sites in a nucleotide sequence; analyzing the nucleotide sequence and available cut sites with a computational model to identify the optimal cut site for introducing the desired genetic change into the nucleotide sequence; and contacting the nucleotide sequence with a DSB-inducing genome editing system, thereby introducing the desired genetic change in the nucleotide sequence at the cut site.

The invention provides preparation of an anti-p-Tau217 antibody and application of the anti-p-Tau217 antibody in an Alzheimer's disease detection kit, and belongs to the technical field of antibodies and immunodetection, the anti-p-Tau217 antibody comprises a heavy chain variable region and a light chain variable region, a coding gene of the heavy chain variable region comprises a nucleotide sequence shown in SEQ ID NO.1, and a coding gene of the light chain variable region comprises a nucleotide sequence shown in SEQ ID NO.2. The coding gene of the light chain variable region comprises a nucleotide sequence shown in SEQIDNO.3, the heavy chain variable region comprises an amino acid sequence shown in SEQIDNO.2, and the light chain variable region comprises an amino acid sequence shown in SEQIDNO.3. The invention also discloses a preparation method. The invention provides preparation of an anti-p-Tau217 antibody and application of the anti-p-Tau217 antibody in an Alzheimer's disease detection kit. The successful expression of the monoclonal anti-p-Tau217 antibody with high affinity is realized, and the monoclonal anti-p-Tau217 antibody is used for detecting the Alzheimer's disease.

The invention discloses an application of a rape BnC05ERF378 gene in improving the waterlogging resistance of a plant. The nucleotide sequence of the rape BnC05ERF378 gene is as shown in SEQ ID NO. 1. According to the invention, a brand new stain-resistant gene BnC05ERF378 is cloned from rape for the first time, and is transferred into arabidopsis thaliana to construct a pure line plant and complete stain resistance identification. Experimental results show that the growth state of the transgenic BnC05ERF378 gene arabidopsis thaliana is remarkably superior to that of a wild type after waterlogging treatment, the transgenic BnC05ERF378 gene arabidopsis thaliana can normally bolt, the survival rate is increased from 12.5% to 37.5%, it is fully proved that the gene can enhance the survivability of the plant in a waterlogging environment by regulating the response mechanism of the plant to waterlogging stress, and the survival rate of the plant is increased from 12.5% to 37.5%. And a key guarantee is provided for stable growth of plants under high-humidity, waterflooding and other adverse conditions. According to the discovery and application of the waterlogging-resistant gene BnC05ERF378 disclosed by the invention, a clear functional gene target is provided for improvement of waterlogging resistance of crops. By constructing an expression vector containing the gene and combining mature technologies such as agrobacterium transformation, the gene can be stably introduced into a target plant, and a pure transgenic plant is obtained.

The invention provides a plant salt tolerance related transcription factor TaVOZ1 as well as a coding gene and application thereof, and belongs to the technical field of functional genes. The TaVOZ1 is related to plant salt tolerance and has an amino acid sequence as shown in SEQ ID No. 1, and a gene for coding the protein has a nucleotide sequence as shown in SEQ ID No. 2. The plant salt tolerance can be regulated by regulating the content or activity of the protein or the expression quantity of the gene, for example, when the gene is over-expressed, the plant salt tolerance is enhanced, and when the gene is knocked down, the salt tolerance is weakened. The TaVOZ1 disclosed by the invention can be combined with 5 '-CTTCT-3'or a reverse complementary sequence 5'-AAGAAG-3 'in a downstream target gene promoter to activate the expression of a stress response gene so as to improve the salt tolerance of a plant, so that the TaVOZ1 can be used for culturing a salt-tolerant wheat variety.

The invention belongs to the technical field of genetic engineering, and discloses a phytosusceptibility-related TaCNGC13 gene, a protein and application of the phytosusceptibility-related TaCNGC13 gene and the protein. The phytosusceptible related TaCNGC13 gene is derived from a wheat variety water source 11, the nucleotide sequence of the gene is shown as SEQ ID NO: 1, and the amino acid sequence of the gene encoding protein is shown as SEQ ID NO: 2. The plant-susceptibility-related TaCNGC13 gene is induced to express by the puccinia striiformis, the expression of the plant-susceptibility-related TaCNGC13 gene in wheat is stably silenced, and the resistance of the obtained transgenic wheat to a plurality of physiological races of the puccinia striiformis is higher than that of wild wheat. The invention provides a novel germplasm material for green control of wheat stripe rust and cultivation of wheat stripe rust resistant varieties.

The invention discloses a nested PCR (Polymerase Chain Reaction) amplification primer for an internally-running sheep disease virus, which comprises a first-round upstream primer, a first-round downstream primer, a second-round upstream primer and a second-round downstream primer, the nucleotide sequence of the first-round upstream primer is as shown in SEQ ID NO: 1, the nucleotide sequence of the second-round upstream primer is as shown in SEQ ID NO: 3, and the nucleotide sequence of the second-round downstream primer is as shown in SEQ ID NO: 4. The nucleotide sequences of the first round of downstream primer and the second round of downstream primer are as shown in SEQ ID NO: 2. The kit has the advantages of excellent specificity and sensitivity, low cost and good stability.

The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

The invention belongs to the technical field of genetic breeding, and relates to an InDel marker for identifying green Chinese onion cytoplasm fertility based on a chloroplast genome and application of the InDel marker. The InDel marker is subjected to amplification detection by adopting a primer group, and the primer group comprises one or more of a first primer group, a second primer group, a third primer group and a fifth primer group; a pair of nucleotide sequences of the first primer group are as shown in SEQ ID NO.1-2; a pair of nucleotide sequences of the second primer group are as shown in SEQ ID NO.3-4; a pair of nucleotide sequences of the third primer group are as shown in SEQ ID NO.5-6; a pair of nucleotide sequences of the fourth primer group are as shown in SEQ ID NO.7-8; and a pair of nucleotide sequences of the fifth primer group are as shown in SEQ ID NO.9 to SEQ ID NO.10. The InDel marker provided by the invention can be used for identifying the cytoplasm fertility of the green Chinese onion, and the tedious procedure and process for extracting DNA can be omitted in the detection process, so that the detection process is simplified.