48 results about "Complementary deoxyribonucleic acid" patented technology

The invention discloses preparation of a human CD56 immune phage display antibody library, which comprises the following steps: S1, animal immunization: expressing recombinant hCD56 protein in escherichia coli, then immunizing a Balb / c mouse, and detecting immune titer; s2, splenocyte preparation: extracting RNA (Ribonucleic Acid) of immune mouse spleen cells, and performing reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid); s3, preparation of an initial bacteriophage display library: amplifying a heavy chain variable region and a light chain variable region by using a PCR technology, and then assembling to form scFv; constructing scFv phagocytids, electrically transforming the scFv phagocytids into TG1 competent cells, and constructing a bacterial library; s4, constructing a phage single-chain library of the hCD56: infecting the bacterial library by using a helper phage M13K07, and constructing a phage single-chain antibody library; and S5, elutriation and screening of a phage library: specifically enriching a phage single-chain antibody library, screening positive clones from the phage single-chain antibody library, and determining a sequence. According to the invention, the human hCD56 antibody scFv sequence fragment can be screened out, and the research and development period is shortened.

The invention belongs to the technical field of microbiological detection, and discloses a composite probe for detecting salmonella enteritidis and a preparation method. The composite probe for detecting the salmonella enteritidis comprises magnetic nanoparticles FeO, wherein the surface of the magnetic nanoparticles FeO is modified with an aptamer apt which is specifically combined with the salmonella enteritidis; the surface of the long afterglow nano particle PLNPs is modified with a DNA (Deoxyribose Nucleic Acid) sequence cDNA (Complementary Deoxyribonucleic Acid) complementary with the aptamer; the aptamer apt and the complementary DNA sequence cDNA are subjected to hybridization to form a composite probe structure FeO-SEapt (at) PLNPs-cDNA (complementary deoxyribonucleic acid). The long afterglow luminescence characteristic of PLNPs (ZnGaO: Cr) is utilized, and an excitation light source is stopped before detection, so that signal acquisition completely avoids autofluorescence of a sample matrix, background fluorescence interference is thoroughly eliminated, and the signal-to-noise ratio is remarkably improved.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-CD38 nano antibody as well as a preparation method and application thereof. The method comprises the following steps: constructing and expressing CD38 antibody protein, immunizing alpaca, obtaining immune alpaca PBMC (peripheral blood mononuclear cells), extracting RNA (ribonucleic acid) of the cells, carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), carrying out PCR (polymerase chain reaction) to obtain antibody gene segments, constructing the antibody gene segments on a phage expression vector to construct a CD38 immune antibody library, carrying out two rounds of panning, carrying out Clone ELISA (enzyme-linked immuno sorbent assay) detection on clones to obtain an antibody combined with the CD38 protein, carrying out sequencing analysis, and selecting a unique sequence. High-throughput expression is carried out through a mammalian cell high-throughput expression system, antibody protein is obtained, FACS detection is carried out through a CD38 high-expression cell line, and finally the antibody combined with CD38 is obtained. The method is simple and rapid to operate, a mammalian cell expression system induces efficient expression of the antibody, processing modification can be performed after translation, and the activity of the antibody is closer to that of a natural antibody.

The invention provides a glass nanopore probe based on an aptamer and application of the glass nanopore probe in dopamine detection, and belongs to the technical field of electrochemical sensing and scanning probe microscopy. The sensor comprises: a glass nanopore probe, the inner wall of which is modified with a gold layer; a cDNA (complementary deoxyribonucleic acid) single chain is fixed on the gold layer through a gold-sulfur bond; a part of the sequence of the dopamine aptamer is hybridized and combined with the cDNA single chain; wherein the combination of the dopamine and the dopamine aptamer causes the change of an ion current rectification signal of the glass nanopore probe, so that the detection of the dopamine is realized based on the change of the ion current rectification signal of the glass nanopore probe. The invention provides a valuable scheme for developing an aptamer nanopore scanning electrochemical sensor for single cell analysis and researching neurotransmitter-related diseases.

The invention discloses an artificially optimized cDNA (complementary deoxyribonucleic acid)-based high-expression trametes versicolor chitin endonuclease gene, a recombinant vector, recombinant bacteria, recombinant enzyme and a preparation method thereof. The optimized trametes versicolor chitin endonuclease gene, such as a sequence as shown in SEQ ID NO.1, is constructed into a pichia pastoris expression vector and then converted into a pichia pastoris expression strain; according to the method, recombinant bacteria expressing active trametes versicolor chitin endonuclease are obtained through high-Zeocin-resistance YPD plate screening, high-expression pichia pastoris transformants are further screened from high-Zeocin-resistance transformants, finally fermentation is carried out in a bioreactor, obtained supernate contains a large amount of recombinant active trametes versicolor chitin endonuclease, and the recombinant active trametes versicolor chitin endonuclease can be obtained under the high-density fermentation condition. The expression level of the recombinant trametes versicolor chitin endonuclease reaches 3.1 g / L, and the trametes versicolor chitin endonuclease is efficiently purified through ammonia sulfate fractional precipitation and ion exchange. The recombinase can hydrolyze colloidal chitin to generate chitin oligosaccharide, and has important application value and wide application prospect in the aspects of preparation of functional chitosan oligosaccharide, antibiosis and anti-tumor, agricultural and biological control, environmental protection, industrial catalysis and the like.

Disclosed herein are methods for processing a cell-free sample of a subject, comprising, providing said cell-free sample of said subject, wherein said cell-free sample comprises a plurality of nucleic acid molecules, subjecting said plurality of nucleic acid molecules to one or more amplification reactions to generate a plurality of complementary deoxyribonucleic acid (cDNA) molecules or derivatives thereof, and sequencing said plurality of cDNA molecules or derivatives thereof. Further disclosed herein are systems, comprising a processor, and a non-transitory computer readable storage medium encoded with a computer program that causes said processor to provide a cell-free sample of a subject, wherein said cell-free sample comprises a plurality of nucleic acid molecules, subject said plurality of nucleic acid molecules to one or more amplification reactions to generate a plurality of complementary deoxyribonucleic acid (cDNA) molecules or derivatives thereof, and sequence said plurality of cDNA molecules or derivatives thereof.

The invention relates to the technical field of biological research data analysis, and discloses a computational biological research data analysis display method, which comprises the following steps of: extracting total RNA (Ribonucleic Acid) from biological tissues or cells, reversely transcribing the RNA into cDNA (Complementary Deoxyribonucleic Acid), constructing a library, and sequencing the library by utilizing a high-throughput sequencing platform to obtain a large number of short sequences. According to the method, the number of the short sequences mapped to the same genome coordinate interval is compared and the threshold value is judged, so that the abnormal repeated area can be effectively identified, whether the gene expression difference result is credible or not is clearly judged, and when the number of the abnormal sequences exceeds the preset repeated sequence threshold value, the influence operation of the repeated sequences is eliminated, and the accuracy of the gene expression difference result is improved. That is to say, completely identical and overlapped short sequences are removed under the condition that the FPKM value is relatively high, and the updating result is compared again, so that the problem of expression quantity distortion caused by repeated reckoning can be corrected.

The invention relates to a double-signal aptamer sensor and a preparation method and application thereof, and belongs to the technical field of sensors. The dual-signal aptamer sensor comprises a first compound, a second compound and a third compound, wherein the first compound comprises Au dimers and functionalized cDNA (complementary deoxyribonucleic acid) modified on the Au dimers; the first compound is Fe3O4 (at) SiO2-CDs, and the second compound is Fe3O4 (at) SiO2-CDs and an aptamer apt of aflatoxin B1, wherein the aptamer apt of aflatoxin B1 is modified on the Fe3O4 (at) SiO2-CDs; the Fe3O4 (at) SiO2-CDs comprise Fe3O4 (at) SiO2 and CDs which are modified on the Fe3O4 (at) SiO2; the first compound and the second compound are connected through complementary base pairing to form an assembly. Due to the introduction of the Raman signal probe, the Raman signal of the assembly is enhanced, meanwhile, the Au dimers have a quenching effect on the fluorescence of the magnetic substrate, and the fluorescence signal of the assembly is weakened. After the target is added, the aptamer is preferentially specifically combined with the target, the assembly is disassembled, the Au dimers fall off from the Fe3O4 (at) SiO2-CDs, and through magnetic separation, a Raman signal of precipitation is reduced along with the addition of the target, and a fluorescence signal is recovered. The dual-mode detection is realized by measuring the change of Raman and fluorescence signals of the precipitation part along with the target addition amount.

The invention discloses a rapid sterile detection method and kit based on a 16S / 18S rRNA-cDNA digital PCR technology, and the method comprises the following steps: S1, extracting total RNA in a to-be-detected sample to obtain bacteria 16S rRNA and fungi 18S rRNA; s2, carrying out reverse transcription on the bacterium 16S rRNA and the fungus 18S rRNA to obtain a stable cDNA (complementary deoxyribonucleic acid) fragment; and S3, carrying out digital PCR amplification by taking the cDNA fragment as a template, and judging whether bacterial contamination exists or not according to an amplification result so as to realize rapid sterile detection. According to the characteristics of high conservative and high copy of bacteria 16S rRNA / fungi 18S rRNA, 30 sets of universal primer probe combinations are verified, and gt can be covered by selecting the optimal two sets of primer probe combinations; 99.99% of common pollution bacteria / fungi in clinic and pharmacy, and no cross amplification with mitochondrial rRNA of mammals such as human, hamster, cattle and the like.

The invention discloses a kit and a method for preparing cDNA (complementary deoxyribonucleic acid) by reverse recording of a single B cell of a mouse. The kit comprises a B cell lysis solution and a reverse transcription buffer solution system, wherein the B cell lysis solution is prepared from a nonionic detergent, dNTP (deoxyribonucleoside triphosphate), dCTP (deoxycytidine triphosphate), a secondary structure stabilizer, a reducing agent, BioIS-mCH2-RT (reverse transcriptase), BioIS-CK-RT, a mouse RNA (Ribonucleic Acid) enzyme inhibitor and nuclease-free water; the reverse transcription buffer solution system comprises 5X RT Buffer, a secondary structure stabilizer, Mg < 2 + >, a reducing agent, a mouse RNA enzyme inhibitor, a Bio-Adaptor primer, reverse transcriptase and nuclease-free water. The kit can significantly improve the success rate, accuracy and sensitivity of reverse transcription of a single B cell, and simplify the operation process.

The invention discloses a polypeptide asparagine ligase VdiPAL1 and an application of the polypeptide asparagine ligase VdiPAL1. The polypeptide asparagine ligase VdiPAL1 is newly found from viola plants, 12 mg / L zymogen protein is obtained through expression of an escherichia coli prokaryotic system, and compared with homologous enzymes VyPAL2 and VdiPAL1 which are widely applied at present, the polypeptide asparagine ligase VdiPAL1 has the advantages of high expression, high catalytic efficiency and substrate specificity and has good development and application potential. The invention also shows the value of the VdiPAL1 for guiding homologous enzyme modification, and particularly discloses the mutant protein VyOPT of the polypeptide asparagine ligase VyPAL2 and a coding cDNA (complementary deoxyribonucleic acid) sequence of the mutant protein VyOPT. The invention also discloses a mutant VyOPT, which comprises five mutations obtained by modification guided by the VdiPAL1. Compared with the expression quantity of the natural VyPAL2, which is about 0.1 mg / L, the expression quantity of the VyOPT reaches 12 mg / L.

The invention relates to the technical field of biological detection, in particular to a method for detecting ECSIT by adopting a fluorescent probe. The ECSIT gene is located in a No.19 chromosome-NC000019.10 of a wisdom, and the nucleotide sequence of the ECSIT gene is as shown in SEQ ID NO: 1; the nucleotide sequences of the PCR amplification primer of the ECSIT are shown as SEQ ID NO: 2 and SEQ ID NO: 3, the primer shown as SEQ ID NO: 2 is a forward PCR primer for amplifying cDNA (complementary deoxyribonucleic acid) corresponding to the ECSIT, and the primer shown as SEQ ID NO: 3 is a reverse PCR primer for amplifying cDNA corresponding to the ECSIT; the nucleotide sequence of the detection probe of cDNA corresponding to ECSIT is shown as SEQ ID NO.4, the 5'end of the probe is marked with a fluorescence reporter group FAM, and the 3 'end of the probe is marked with a fluorescence quenching group MGB. The detection probe has the advantages of high sensitivity, high accuracy and non-invasiveness, and is a safe and reliable gene detection means.

The improved nucleic acid capturing method comprises the following steps: hybridizing a random probe and an oligonucleotide chain fixed on a capturing chip through a splint oligonucleotide, reversely transcribing RNA (Ribonucleic Acid) in a sample captured by the random probe into cDNA (Complementary Deoxyribonucleic Acid), connecting the cDNA to the oligonucleotide chain, and carrying out reverse transcription and connection in the same reaction system. In the process, RNA is subjected to reverse transcription to form cDNA, and meanwhile, the cDNA is connected to an oligonucleotide chain fixed on a chip by virtue of fixed oligonucleotide sequences at two ends hybridized by splint. Through the method, the number of captured genes can be increased, the loss of cDNA (complementary deoxyribonucleic acid) is reduced, the original three steps are combined into one step, and the duration of the process can be further shortened.

The invention relates to the technical field of biological genetic engineering, in particular to a wheat salt-tolerant nuclear factor Y subunit gene TaNF-YC8 and application thereof. The cDNA (complementary deoxyribonucleic acid) sequence of the TaNF-YC8 gene is as shown in SEQ ID NO. 1; according to the application, by silencing the TaNF-YC8 gene or inhibiting the expression activity of the TaNF-YC8 gene, the resistance of wheat to salt stress can be promoted, and the application has extremely important practical significance for cultivating new wheat varieties with higher salt resistance and improving the yield and quality of wheat in a stress environment.

The invention discloses a tobacco cytochrome c1 subunit and application thereof in plant disease resistance. Belongs to the technical field of gene engineering. The cDNA (complementary deoxyribonucleic acid) nucleotide sequences of the tobacco NtCytc1-1a / 1-1b / 1-2a / 1-2b / 1-2c / 1-2d gene are as shown in SEQ ID NO: 1-SEQ ID NO: 6. The function loss of the protein coded by the tobacco NtCytc1-1a / 1-1b / 1-2a / 1-2b / 1-2c / 1-2d gene leads to mROS outbreak, disease resistance enhancement and GmMEKK1 response enhancement of the plant. The method has a guiding effect on cultivation of new disease-resistant varieties of crops.

The invention belongs to the technical field of biomaterial chemistry, and particularly discloses a preparation method and application of functional escherichia coli for expressing laccase. The method comprises the following steps: firstly, according to a full-length cDNA (complementary deoxyribonucleic acid) sequence of a laccase gene, integrating a cotA target gene into a vector plasmid and constructing a recombinant expression plasmid; and introducing the recombinant expression plasmid into escherichia coli to prepare functionalized recombinant escherichia coli. Wherein the active center of the laccase contains copper ions [Cu (II)]. The binding domain of the laccase is mainly composed of histidine, cysteine, methionine and the like, and the laccase can utilize molecular oxygen as an electron acceptor to oxidize various phenolic and non-phenolic compounds and increase active sites except cytochrome c in escherichia coli. Meanwhile, when the substrate is oxidized, the method has the advantages of substrate specificity, mild reaction conditions, harmlessness to the environment and the like, molecular oxygen in air is used as an electron acceptor, only water is generated as a byproduct, the principle of environmental protection is met, and the oxygen reduction capacity of the microbial fuel cell is accelerated.

The invention relates to the field of plant molecular biology, in particular to application of a CsERF113L gene or a biological material containing the CsERF113L gene in regulation and control of stress resistance and yield of cucumbers. The cDNA (complementary deoxyribonucleic acid) sequence of the CsERF113L gene is as shown in SEQ ID NO. 1. The CsERF113L gene is knocked out in cucumber by adopting a CRISPR-Cas9 gene editing technology, the antioxidant capacity and photosynthetic capacity of cucumber plants are remarkably enhanced, the saline-alkaline resistance is remarkably improved, the root growth is accelerated, the cucumber yield is remarkably increased, and knockout strains and over-expression strains have opposite trends. The result shows that the CsERF113L gene affects the regulation and control mechanism of cucumber stress resistance. The CsERF113L gene has important guiding significance and application value for guiding widening of cucumber cultivation conditions in actual production, improvement of cucumber high yield and stress resistance and cucumber seed selection and breeding.

The invention belongs to the technical field of insecticides, and particularly relates to a ds-ABC transporter gene for preventing and controlling resistant Frankliniella occidentalis as well as a screening method and application of the ds-ABC transporter gene. The ABC transport protein gene is selected from any one or more of ABCG20, ABCA1, ABCB8, ABCD2 and ABCA5, and the ABC transport protein gene is selected from any one or more of ABCG20, ABCA1, ABCB8, ABCD2 and ABCA5. The screening method comprises the following steps: (1) testing the resistance level of the Frankliniella occidentalis to spinetoram, imidacloprid and thiamethoxam; (2) extracting and purifying the total RNA of the Frankliniella occidentalis population by using a trace total RNA extraction kit, synthesizing cDNA (complementary deoxyribonucleic acid) by taking the RNA as a template, and then carrying out quantitative primer design and gene expression; and (3) carrying out significant analysis on difference analysis of all data through real-time fluorescent quantitative detection, and screening out 5 ds-ABC transporter genes which are subjected to significant up-regulation expression in a resistant population. According to the invention, the key ABC transporter gene for mediating the resistance of the Frankliniella occidentalis to the spinetoram is screened and determined, and a basis is provided for development of a novel anti-resistance insecticide taking the ABC transporter as a target.

The invention discloses an oligonucleotide pharmacobiological analysis system based on a GalNAc technology, and relates to the technical field of drug analysis, and the oligonucleotide pharmacobiological analysis system is technically characterized in that in a GalNAc conjugate coupling delivery module, a coupling unit covalently couples an oligonucleotide drug and a GalNAc-trimer through a click chemical reaction to form a liver-targeted delivery carrier; the targeting unit is specifically combined with an asialoglycoprotein receptor on the surface of a hepatocyte by utilizing the vector to realize endocytosis; in the RT-qPCR quantitative analysis module, a magnetic separation purification unit adsorbs oligonucleotides by using carboxylated magnetic beads; the reverse transcription unit uses a stem-loop primer to convert oligonucleotide into cDNA (complementary deoxyribonucleic acid) under the action of heat-resistant reverse transcriptase; the fluorescence quantification unit adopts an FAM-labeled TaqMan probe to monitor amplification, and the absolute concentration of the medicine is calculated in combination with a standard curve; according to the invention, the technical problems of poor targeting, serious off-target effect and poor stability are solved, and high-quality drug effect evaluation can be carried out.

The invention relates to the technical field of plant genetic engineering, in particular to a gene down-regulation method based on regulation of precursor mRNA (messenger Ribonucleic Acid) splicing efficiency, which comprises the following steps: firstly, identifying at least one intron 5'terminal splicing site containing a GT sequence in a target gene; aiming at the site, designing a guide RNA (Ribonucleic Acid) sequence capable of targeting the downstream fourth or fifth nucleotide, and verifying the cleavage activity of the guide RNA sequence; constructing a gene editing vector by using the guide RNA sequence which is verified to be effective and a Cas9 nuclease coding gene; introducing the vector into a plant cell, carrying out genetic transformation and regeneration to obtain a transformed plant, and screening out a mutant which is subjected to site-directed editing at a target site through DNA (Deoxyribose Nucleic Acid) sequencing; and finally, extracting RNA (Ribonucleic Acid) of the mutant, reversely transcribing into cDNA (Complementary Deoxyribonucleic Acid), and analyzing transcript change through PCR (Polymerase Chain Reaction) amplification and gel electrophoresis. According to the invention, stable, heritable and accurate knock-down of gene expression is realized, and a new tool is provided for essential gene function research and crop character improvement.

The invention relates to the technical field of biological genetic engineering, in particular to a wheat salt-tolerant DNA damage repair gene TaSOG1 and application thereof. The cDNA (complementary deoxyribonucleic acid) sequence of the TaSOG1 gene is as shown in SEQ ID NO. 1; according to the application, the resistance of wheat to salt stress is promoted by silencing the TaSOG1 gene or inhibiting the expression activity of the TaSOG1 gene. The method has extremely important practical significance for cultivating new wheat varieties with stronger salt resistance and improving the yield and quality of the wheat in a stress environment.

The invention discloses a cotton gene GhFBH1 and application thereof in identifying the height of a first fruit branch of cotton, and belongs to the field of biotechnology application. The cDNA (complementary deoxyribonucleic acid) sequence of the gene GhFBH1 associated with the first fruit branch height character of the cotton is as shown in SEQ ID NO. 1, and the genome sequence is as shown in SEQ ID NO. 2. The GhFBH1 is regulated and controlled by an SNP site on the gene, and after the SNP site is mutated, the height of a first fruit branch is obviously lower than that of a wild type. The gene has important research value and application prospect in efficient identification of cotton varieties with low first fruit branch height and improvement and breeding of excellent cotton varieties.

The invention discloses an identification method and application of a larimichthys crocea immortalized cell line. The identification method comprises the following steps: 1, providing a larimichthys crocea cell sample; step 2, screening the larimichthys crocea cell immortalized genes, namely extracting total RNA (Ribonucleic Acid) of each cell sample, reversely transcribing the total RNA into cDNA (Complementary Deoxyribonucleic Acid), measuring the expression quantity of immortalized related genes, and determining the genes with the expression quantity conforming to immortalized index characteristics, namely the larimichthys crocea cell immortalized genes; and step 3, based on the larimichthys crocea cell immortalized gene, identifying whether the to-be-identified larimichthys crocea cell is an immortalized cell. The method for identifying the immortalized cells of the pseudosciaena crocea greatly shortens the traditional time dependent on long-term passage, avoids the risk of noise pollution in long-term culture, improves the probability of successful identification, and has the characteristics of low cost and high efficiency.

The invention discloses a gibberellic disease resistance related receptor protein kinase gene as well as an expression vector and application thereof, and belongs to the field of gene engineering. The cDNA (complementary deoxyribonucleic acid) sequence of the TaRLK3-3B is SEQ ID NO. 1, and the amino acid sequence coded by the TaRLK3-3B is SEQ ID NO. 2. The gene is inserted into an expression vector pAHC25 to obtain an overexpression vector of the gene, and the overexpression vector of the gene is introduced into an infected wheat variety, so that the resistance of the wheat variety infected with the gibberellic disease to the gibberellic disease can be remarkably improved. The TaRLK3-3B is used for genetic engineering breeding, and when the TaRLK3-3B is introduced into wheat varieties susceptible to gibberellic disease, the gibberellic disease resistance of wheat can be improved.

The invention belongs to the technical field of biotechnology, and particularly relates to a construction method of a cDNA (complementary deoxyribonucleic acid) library for screening drought-resistant genes of black-heart golden chrysanthemum. The method disclosed by the invention is scientific and reasonable in design, and the pYES2-NTB no-load transformation yeast strain INVSc1 is firstly used as a control group; secondly, transforming recipient bacteria INVSc1 by using the constructed black-heart golden chrysanthemum library, and preparing a yeast working bacteria solution; and then, screening by using SG-U liquid culture mediums with different concentrations of PEG 3350, after determining the screening concentration, coating the library working bacterial liquid on an SG-Ura plate for library positive clone identification, and sequencing the positive clone at the same time. Finally, rotation verification is carried out; according to the method, the drought-resistant gene of the black-heart golden chrysanthemum is determined through yeast screening, compared with the method that the drought-resistant gene of the black-heart golden chrysanthemum is identified after genetic transformation of species, the yeast screening period is short, the efficiency is high, and the species do not have a genetic transformation technology.

The invention discloses a nornicotine negative regulation and control gene and cloning and application thereof, tobacco and a tobacco product, and belongs to the technical field of genetic engineering, the nornicotine negative regulation and control gene is a tobacco NtWRKY51 gene, and can negatively regulate and control nornicotine in tobacco. The cloning method of the nornicotine negative regulation nornicotine gene comprises the following steps: designing a specific primer, amplifying full-length cDNA (complementary deoxyribonucleic acid) and a genome sequence, and realizing PCR (polymerase chain reaction) amplification; and purifying the PCR product. The overexpression vector of the nornicotine negative regulation nornicotine gene is pCHF3. The application of the nornicotine negative regulation and control gene is realized by negatively regulating and controlling accumulation of nornicotine in tobacco leaves by inhibiting expression of CYP82E4 through the nornicotine negative regulation and control gene. The tobacco plant is formed by breeding transgenic tobacco prepared by applying the nornicotine negative regulation nornicotine gene. Through overexpression of the NtWRKY51, the nornicotine content of the plant can be reduced by 40% to the maximum, and the NtWRKY51 has a great application prospect in the aspect of tobacco with low nornicotine content.

The invention discloses a gene for increasing sorghum yield and application thereof, and belongs to the field of plant genetic engineering. The cDNA (complementary deoxyribonucleic acid) sequence of the SbWRKY50 (Sobic.003G037400.1) is equal to '1', and the amino acid sequence coded by the SbWRKY50 (Sobic.003G037400.1) is equal to '2'. The gene is from sorghum, and overexpression of SbWRKY50 in sorghum can delay leaf senescence and increase the yield of sorghum. The gene leaf is knocked out through a CRISPR / Cas9 technology to generate a phenotype which is aged in advance, so that the yield of sorghum is reduced. Therefore, the SbWRKY50 is a gene for increasing the yield of sorghum and is expected to be used for genetic engineering breeding.

The invention discloses a gene associated with cotton fiber length and application, and belongs to the field of biotechnology application. The cDNA (complementary deoxyribonucleic acid) sequence of the gene GhDTX48 associated with the cotton fiber length character is shown as SEQ ID NO. 1, and the genome sequence is shown as SEQ ID NO. 2. The gene GhDTX48 is regulated and controlled by an adjacent SNP site (D11: 24631774), the SNP basic group is mutated from T to C, and the length of genotype fiber after mutation is obviously greater than that before mutation. The invention further provides a method for identifying the variety of the long-fiber cotton, and the cotton with the basic group C corresponding to the SNP site is the variety of the long-fiber cotton. Meanwhile, a new variety of cotton long fibers is cultivated through a genetic engineering means, a gene containing a high-yield genotype GhDTX48 (C) is transferred into a cotton variety, or an SNP site in a low-yield genotype GhDTX48 (T) is subjected to site-specific mutagenesis to form (C), so that the new variety of cotton long fibers is cultivated. The gene has important research value and application prospect in efficient identification of long-fiber cotton varieties and improvement and breeding of excellent cotton varieties.

The invention relates to the technical field of gene diagnosis, and particularly discloses a liver cancer diagnostic kit based on a GBP2 gene and a detection method, the kit comprises an RNA extraction component used for extracting total RNA from a liver cancer or para-carcinoma tissue sample to obtain an RNA sample; the reverse transcription component is used for carrying out reverse transcription reaction on the RNA sample to obtain cDNA (complementary deoxyribonucleic acid); carrying out RT-qPCR detection on the components; the fluorescent dye premix liquid is used for carrying out real-time quantitative PCR amplification on the cDNA to obtain a GBP2 gene expression Ct (GBP2) value; the internal reference gene detection component comprises a GAPDH primer pair and is used for obtaining an internal reference Ct (GAPDH) value; the components are interpreted, delta Ct is obtained through calculation, a preset threshold value is compared with delta Ct, and the GBP2 expression level is obtained through judgment; the GBP2 is used as a liver cancer specific diagnosis marker, the detection rate of early liver cancer can be improved by detecting the expression level (such as mRNA or protein level) of the GBP2, and the gene is particularly suitable for AFP negative or low expression patients and has higher sensitivity and specificity.

The invention discloses a dendrobium officinale cellulose-like synthase protein as well as a coding gene and application thereof, and relates to the technical field of biochemistry. The nucleotide sequence of the coding gene of the protein is as shown in SEQ ID No. 1; the cloning method comprises the following steps: culturing dendrobium officinale seedlings for two weeks, sampling, and extracting total RNA (Ribonucleic Acid); synthesizing cDNA (complementary deoxyribonucleic acid) through reverse transcription; designing a primer by taking the primer as a template, and carrying out PCR (Polymerase Chain Reaction) amplification; after gel electrophoresis of the amplified fragment is finished, recovering a product, connecting the product to a cloning vector pMD18-T, transforming escherichia coli DH5alpha, and sequencing after identification; the gene is used for transforming arabidopsis thaliana and yeast; the gene can be applied to directionally breeding new varieties of low-temperature-resistant dendrobium nobile. The invention has important significance for improving the low-temperature resistance of plants and cultivating high-quality and high-resistance new varieties.