42 results about "Nucleotide sequencing" patented technology

The invention belongs to the technical field of genetic engineering, in particular to specific molecular markers of sex chromosomes of Tilapia nilotica, wherein the nucleotide sequences of specific molecular markers NtY1 and NtY2 of a Y chromosome are respectively shown in SEQID No.1 and SEQID No.2, and the nucleotide sequence of a specific molecular marker NtX1 of a X chromosome is shown in SEQID No.3; in the invention, a genetic sex identification method of Tilapia nilotica is established by using the specific molecular markers of the sex chromosomes of the Nile tilapia, and through carrying out PCR amplification on any one or more than one of the specific molecular markers of the sex chromosomes, YY super-male fishes, XY milter and XX female fishes can be identified economically, quickly and accurately, thus being beneficial to realizing the large-scale production of holandric Tilapia nilotica fries through YY super-male fish breeding routes, obviously improving the breeding production of Tilapia nilotica, reducing the breeding cost, and improving the economic benefit.

The invention belongs to the technical field of animal infectious disease prevention and curing, and particularly relates to application of mycoplasma bovis MbovP730 protein in natural infection and vaccine immunity identification. According to the codon preference of escherichia coli, the Mbov_730 gene of M.bovis HB0801 is cloned and is modified, and a mycoplasma bovis tryptophan codon UGA is mutated into a tryptophan codon UGG in the escherichia coli so as to express recombinant mycoplasma bovis MbovP730 protein in the escherichia coli. The nucleotide sequence of the MbovP730 protein is disclosed in SEQ ID NO: 13, and the protein can be used as the antigen protein to be applied to the preparation of a kit for identifying mycoplasma bovis wild strain infection and vaccine strain immune identification.

The invention specifically discloses a tea tree MYB transcription factor CsAN1 and an application thereof in regulation of anthocyanin metabolism. A nucleotide sequence of the tea tree MYB transcription factor CsAN1 is represented as SEQ ID NO:1. CsAN1 is connected with a 35S promoter and then is connected to a pBI121 carrier, meditated transformation of tobaccos is realized by agrobacteria GV3101, meanwhile, the gene as well as genes CsGL3 and CsTTG1 capable of forming compounds with the gene is used for transforming the tobaccos respectively, leaves of overexpressed tobacco plants turn red, anthocyanin content detected by a spectrophotometer is remarkably increased, and two main pigments including cyanidine and delphinidin are detected through HPLC (high performance liquid chromatography).

The invention discloses a CRISPR-Cas12a detection primer group for francisella tularensis and an application thereof. The invention provides the primer group for detecting francisella tularensis and is composed of an RPA primer pair and crRNA; the RPA primer pair is designed according to the nucleotide sequence shown in SEQ ID NO.1 in the francisella tularensis genome; the crRNA comprises an anchoring sequence and a guide sequence, wherein the anchoring sequence can be combined with a cas protein, and the guide sequence is matched with an amplification product sequence of an RPA primer pair. The detection primer group has the characteristics of short detection time, high amplification efficiency, high sensitivity, strong specificity and the like, and the detection sensitivity on francisella tularensis positive plasmids can reach 900 copy/ml, and the detection sensitivity on the francisella tularensis genome can reach 1260 fg/mL. The francisella tularensis onsite diagnosis method has awide application prospect.

The invention belongs to the field of microbial genetic engineering, and discloses a GTP cyclohydrolase I gene folE and application. A nucleotide sequence of the GTP cyclohydrolase I gene folE is shown as SEQ ID NO:1. The gene is derived from food-borne lactobacillus plantarum, is safe and can be used in the field of later-stage food fermentation. A gene knockout strain screening method can be used for improving the screening efficiency of knockout strains. The key role of gene folE gene in folic acid synthesis is proved, and a certain theoretical basis is provided for the research and development of folic acid synthesis functional food. The GTP cyclohydrolase I gene folE and application analyze that the gene is derived from food-grade microorganisms and is safe; and the GTP cyclohydrolaseI gene folE and application analyze that the gene folE gene has nothing to do with cell morphology and strain growth except playing an important role in folic acid synthesis.

The invention discloses an HCV (hepatitis C virus) core antigen, which is a polypeptide encoded by a DNA (deoxyribonucleic acid) of which the amino acid sequence is shown in SEQ ID NO.1 and the nucleotide sequence is shown in SEQ ID NO.2. The invention also discloses a hybridoma cell line secreting monoclonal antibodies, the hybridoma cell line is named as SC16-H6 and has been preserved in the China General Microbiological Culture Collection Center on July 14th, 2011, and the preservation number is CGMCC No. 5074. The invention also discloses another hybridoma cell line secreting monoclonal antibodies, the hybridoma cell line is named as SC23-G5 and has been preserved in the China General Microbiological Culture Collection Center on July 14th, 2011, and the preservation number is CGMCC No. 5075. The invention also discloses a monoclonal antibody of an anti-HCV core antigen, which is secreted by the hybridoma cell line (preservation number: CGMCC No. 5074) or the hybridoma cell line (preservation number: CGMCC No. 5075), and has a specificity to the polypeptide shown in SEQ ID No.1.

The invention discloses an SNP marker related to Erhualian sow litter traits and a detection method and application thereof. The SNP marker is on the nucleotide sequence of the gene of ectonucleoside triphosphate diphosphohydrolase-1 (ENTPD1) of a pig chromosome 14; the site of the SNP marker is the g.117011153 nucleotide site of a chromosome 14 in the reference sequences of International Pig Genomic Sequence of version 10.2 and obtains A/G polymorphism, and the SNP marker is significantly related to the total litter size of Erhualian sow. The invention also discloses a primer pair for detecting the SNP marker, wherein the forward primer is SEQ ID NO: 2, and the reverse primer is SEQ ID NO: 3. The SNP marker is related to the litter performance of the Erhualian sow. Therefore, a high-yield Erhualian sow event can be screened by identifying the SNP marker, and the obtained high-yield Erhualian sow event has substantial economical benefits and social values.

Disclosed is a process for producing the L-form of an amino acid enzymatically from an enantiomeric mixture of the amino acid. The process includes the step of contacting a transformant or a treated product thereof with an enantiomeric mixture of the amino acid. The transformant includes a single host and one or more recombinant vectors introduced thereinto, in which the one or more recombinant vectors include one or more expression vectors and nucleotide sequences integrated thereinto respectively, and the nucleotide sequences are a nucleotide sequence encoding an enzyme capable of converting the D-form of an amino acid into a keto acid and a nucleotide sequence encoding an enzyme capable of converting a keto acid into the L-form of an amino acid. According to this process, a reaction for converting the D-form of an amino acid into a keto acid and a reaction for converting the keto acid into the L-form of the amino acid can be conducted in a single step. Disclosed also is a transformant capable of converting into the L-form of an amino acid as accumulated in a high concentration. The L-form of an amino acid can be efficiently produced from an inexpensive starting material by using the transformant.

The invention discloses a rice secretary type thioredoxin gene, coded proteins thereof and application thereof and aims to provide the rice secretary type thioredoxin gene, the coded proteins thereof and a method for improving antioxidant activities of plants and delaying plant senescence by using the gene. The nucleotide sequence of the rice secretary type thioredoxin gene provided by the invention is a No.1 DNA sequence in a sequence table; and the amino acid sequence coded by the secretary type thioredoxin gene is a No.2 amino acid residue sequence in the sequence table. Experiments show that when the gene is excessively expressed in the rice, hydrogen peroxide content of a plant can be reduced and chlorophyll content can be improved in a normal state or in the presence of salts, so a rice senescence process is delayed. The rice secretary type thioredoxin gene, the coded proteins thereof and the application thereof have important application in aspects of improving the anti-oxidation and anti-aging performance of rice by using molecular breeding.

The invention discloses one group of nucleic acid aptamers capable of specifically bonding with tuberculosis mycobacterium Beijing genotype strain surface lipolysaccharide namely mannose modified mannosylated lipoarabinomannan (ManLAM) and application of the nucleic acid aptamers. The aptamers are specific to small-molecular single-chain DNA (ssDNA) of the ManLAM, and have a nucleotide sequence as shown in the SEQ ID No.1-5. The ssDNA has functions similar to those of a monoclonal antibody, and can be directly and specifically bonded with ManLAM. By adopting an enzyme-linked oligonucleotide assay (ELONA) method, the ManLAM antigen in serum of patients with pulmonary tuberculosis, extrapulmanory tuberculosis and the like can be quickly effectively detected in 2 hours, the ManLAM antigen in sputum of an active pulmonary tuberculosis patient can be directly identified, and the detection sensitivity on the Man LAM antigen can be 0.035mu g/mL. The aptamers are simple to prepare and low in cost, and can be used as an effective novel reagent for diagnosing the mycobacterium tuberculosis antigen of early and latent tuberculosis.

The present invention provides methods for producing a transgenic soybean plant, soybean plant part, or soybean plant cell having increased resistance to soybean cyst nematode parasitism, comprising introducing into the soybean plant, soybean plant part, or soybean plant cell, a heterologous miR164 nucleotide sequence, wherein the heterologous miR164 nucleotide sequence is expressed in the soybean plant, soybean plant part or soybean plant cell, thereby producing a transgenic soybean plant, soybean plant part, or soybean plant cell having increased resistance to soybean cyst nematode parasitism relative to a control soybean plant, soybean plant part, or soybean plant cell. The invention further provides methods of reducing soybean cyst nematode cyst formation in a soybean plant, soybean plant part, or soybean plant cell as well as compositions including nucleic acid constructs for transforming a plant, plant part or plant cell and transformed plants, plant parts and plant cells.

The invention belongs to the technical field of molecular biology and discloses a molecular marker for identifying rice grain length characters, an identification method and application. The molecular marker is GS3-TPAP and comprises GS3-TPAP-O-F with the nucleotide sequence shown as SEQ ID NO.1, GS3-TPAP-O-R with the nucleotide sequence shown as SEQ ID NO.2, GS3-TPAP-I-F with the nucleotide sequence shown as SEQ ID NO.3, and GS3-TPAP-I-R with the nucleotide sequence shown as SEQ ID NO.4. The molecular marker can be used for directly identifying two allelotypes of the rice grain length characters of the GS3 gene, the accuracy reaches 100%, the two allelotypes can be distinguished well, influences of environmental conditions are avoided, and the identifying method is low in consumption and suitable for large-scale application and popularization.

The present invention discloses diamondback moth cecropin 3, a preparation method and an application thereof, wherein the diamondback moth cecropin 3 gene has a nucleotide sequence represented by SEQIDNO:1-2 and has a corresponding amino acid sequence represented by SEQIDNO:3, and sequence analysis results show that the mature peptide of the diamondback moth cecropin 3 has an amino acid sequence represented by SEQIDNO:4. According to the present invention, the diamondback moth cecropin 3 gene or the mature peptide sequence thereof is subjected to prokaryotic expression to obtain the diamondback moth cecropin 3, and the obtained diamondback moth cecropin 3 provides strong inhibition effects for gram-negative bacteria and gram-positive bacteria, provides strong inhibition effects for pathogenic fungi, especially provides strong inhibition effects for a lot of pathogenic fungi of melons and fruits, and is expected to be a novel peptide antibiotic.

The invention belongs to the technical field of biology and in particular relates to an absolute fluorescence quantitative PCR (Polymerase Chain Reaction) primer and a kit for detecting an atypical porcine pestivirus (APPV). Nucleotide sequences of the absolute fluorescence quantitative PCR primer for detecting the atypical porcine pestivirus are shown as SEQ ID NO. 1 and SEQ ID NO. 2; the primer can be used for specifically amplifying an NS3 gene segment of the atypical porcine pestivirus. The invention further provides the kit for detecting the atypical porcine pestivirus and a method for detecting the atypical porcine pestivirus; the kit has the beneficial effects of rapidness and high efficiency, convenience for operation, high specificity, high sensitivity, simplicity for identification, low cost and the like. The primer, the kit and the method, provided by the invention, are used for carrying out quantitative detection and determination on the atypical porcine pestivirus and can be used for rapidly and accurately detecting the copy number of targeting genes and a technological foundation is provided for detecting and identifying the APPV.

The invention belongs to the field of biological medicines, and particularly relates to a kit for detecting human herpes virus infection and a detection method thereof. The invention firstly disclosesa kit for simultaneously detecting multiple herpes viruses, the kit comprises a primer probe system for detecting human herpes virus infection, and the primer probe system comprises a nucleotide sequence group as shown in SEQ ID NO: 1-21. When the kit is used for herpes virus detection, the detection period is shortened to 3-4 hours, and clinically common virus infection can be rapidly, efficiently and accurately determined. The detection method provided by the invention can be used for rapidly monitoring the copy number change of the specific virus in the body of the virus infected patient in real time, evaluating in time and providing adjuvant therapy suggestions for clinicians. Compared with the traditional multiple PCR scheme, the efficient multiple multicolor digital PCR experimentalscheme disclosed by the invention has the advantage that the detection method is more efficient and accurate.

The invention discloses a molecular marker of maize transcription factor ZmPIF3 transgenic rice and application of the molecular marker and belongs to the field of crop genetic breeding. A maize transcription factor ZmPIF3 transgenic rice material which is obtained already has obvious water-saving drought-enduring phenotype. According to a nucleotide sequence of maize transcription factor ZmPIF3 gene, a primer of the molecular marker is designed in CDS, PCR technology is applied to acquire DNA segments of ZmPIF3 and sequence the DNA segments, and the primer of the molecular marker is obtained according to specific segment screening of ZmPIF3. The molecular marker is high in repeatability, stable in amplification, simple and convenient to operate and low in cost and can be used for detecting maize ZmPIF3 genes in rice background, and a solid foundation is laid for molecular marker assisted selection in the future by utilizing ZmPIF3 gene transgenic rice.

The invention provides a single-chain antibody capable of specifically recognizing Reg3A protein, including the nucleotide sequence and amino acid sequence of the heavy and light chain variable regions encoding the protein, and the CDR region determining the specificity of the antibody. In the present invention, a high-affinity single-chain antibody targeting Reg3A protein is screened out through a phage antibody library, and then the Reg3A single-chain antibody is obtained through construction, expression, and purification. The Reg3A single-chain antibody of the present invention can specifically bind to Reg3A, so it can treat or detect diseases related to excessive and abnormal expression of Reg3A.

The invention discloses an efflux pump LexABC participating in self resistance of a phenazine substance myxin as well as a coding gene and application of the efflux pump LexABC. The nucleotide sequence of the efflux pump LexABC is as shown in SEQ ID NO. 1. A lexA gene, a lexB gene and a lexC gene are provided. The genes lexA, lexB and lexC and a vector pBBR1-MSC5 are cloned through seamless connection, so that a recombinant plasmid pBBR-lexABC is obtained. The recombinant plasmid pBBR-lexABC is transformed into antibiotic lysobacter sp. OH13, such that the LexABC overexpressed recombinant strain is obtained. By overexpressing the LexABC protein, the resistance of the myxin-producing strain to the myxin can be enhanced, the myxin-producing strain is protected from toxicity from high-concentration myxin, and important practical significance is provided for culturing a myxin high-yield strain and developing the myxin-producing strain as a biopesticide.

The invention discloses a panax notoginseng reverse osmosis protein gene PnOLP1. The gene PnOLP1 has a nucleotide sequence shown as SEQ ID NO:1, and encodes protein having an amino acid sequence shownas SEQ ID NO:2. It is verified by functional genomics related technical research that the gene PnOLP1 has the function of improving the antifungal property of plants; after the antifungal gene PnOLP1is constructed onto a plant expression vector and transferred into nicotiana tabacum for overexpression, the transgenic nicotiana tabacum plant has quite high in-vitro antifungal activity as a result, and the experimental result shows that the transgenic nicotiana tabacum overexpressing PnOLP1 has a significant inhibiting effect on the growth of four fungi including colletotrichum gloeosporioides, fusarium solani, fusarium oxysporum and nigrospora oryzae.

The invention belongs to the technical field of white spirit detection, and discloses primers, a kit and an identification method for identifying bacillus amyloliquefaciens in yeast for making hard liquor. The primer for identifying the bacillus amyloliquefaciens in the yeast for making hard liquor is a pair of specific ARMS primers: ARMS-1F and ARMS-1R, and the nucleotide sequences of the ARMS-1F and the ARMS-1R are respectively shown as SEQ ID NO: 1 and SEQ ID NO: 2. A qPCR technology is utilized, an ARMS-qPCR detection method for bacillus amyloliquefaciens is established, the bacillus amyloliquefaciens and other related strains in yeast for making hard liquor are distinguished, specificity and detection limit verification shows that the method can be used for rapidly and accurately identifying, and compared with traditional morphology and physiological and biochemical identification means, the method has the advantages that errors are large, and efficiency is low, and the method can be used for rapidly and accurately identifying the bacillus amyloliquefaciens and other related strains in the yeast for making hard liquor for making hard liquor for making hard liquor for making hard liquor for making hard liquor for making hard liquor for making hard liquor for making hard liquor for making hard liquor. The method is simple and convenient to operate, quick in reaction and capable of realizing accurate qualitative identification.