71 results about "Complementary deoxyribonucleic acid" patented technology

The invention belongs to the technical field of biology, and particularly provides a reverse transcriptase UCRT v6 mutant with improved thermal stability, the reverse transcriptase UCRT v6 mutant comprises four single-point mutants, six double mutants and four three-point mutants, and compared with wild type reverse transcriptase UCRT v6, the half-life period of the mutant is longer at 65 DEG C; the three mutants are better in effect, and the half-life period of the three mutants is about 5 times that of the wild reverse transcriptase UCRT v6. The invention also provides a genetically engineered bacterium containing the mutant. The reverse transcriptase UCRT v6 mutant obtained by the construction method provided by the invention has better thermal stability, shows higher thermal stability when being used for reverse transcription of RNA (Ribonucleic Acid) into cDNA (Complementary Deoxyribonucleic Acid) at higher temperature, and has higher application potential.

The invention discloses a Chinese orchid flowering regulation gene CsFTL3 as well as an encoding protein and application thereof, and belongs to the field of plant molecular biology. The gene CsFTL3 for regulating and controlling flowering of Chinese cymbidium is separated from flower bud cDNA (complementary deoxyribonucleic acid) of a Chinese cymbidium variety 'Xiaoxiang', and 174 amino acids are encoded by the gene CsFTL3. Through construction of an expression vector of the CsFTL3 gene, overexpression in wild-type arabidopsis thaliana and heterologous overexpression of the CsFTL3 gene in the arabidopsis thaliana, it is found that bolting and flowering periods of the arabidopsis thaliana are significantly delayed, flower stems are significantly thickened and increased, the number of rosette leaves is significantly increased, and the growth period is prolonged, which indicates that the gene has the function of delaying the flowering time of the arabidopsis thaliana. The Chinese orchid flowering regulation and control gene can be used for molecular mechanism research of orchid flowering time, delay Chinese orchid flowering time, promote plant vegetative growth, prolong whole Chinese orchid viewing time and promote Chinese orchid genetic improvement and variety breeding.

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 gene engineering, and provides a gene OsMYB1 and application thereof in negative regulation of anthocyanin synthesis. The cDNA (complementary deoxyribonucleic acid) sequence of the gene OsMYB1 is as shown in SEQ ID NO. 1. A systematic molecular biology experiment proves that the OsMYB1 inhibits the expression of the OsDFR by combining with a promoter region of an anthocyanin synthesis key structural gene OsDFR, so that the synthesis of anthocyanin is inhibited. Meanwhile, the OsMYB1 can form an MBW ternary complex with the OsB2 and the OsPAC1 to competitively inhibit the synthesis of the anthocyanin, which indicates that the transcription factor OsMYB1 is a negative regulation factor for the synthesis of the rice anthocyanin, the content of the rice anthocyanin can be increased by knocking out the transcription factor OsMYB1, and a certain theoretical basis is provided for cultivating high-quality purple rice with high anthocyanin content.

The invention provides a lentivirus for treating craniopharyngeal tubuloma as well as a preparation method, application and a corresponding pharmaceutical composition of the lentivirus. The lentivirus carries an hTERT (human telomerase reverse transcriptase) promoter and a CDC6shRNA (complementary deoxyribonucleic acid) gene. According to the invention, an hTERT promoter and a CDC6shRNA gene sequence are inserted behind an enzyme cutting site of a vector, an obtained plasmid is transferred into an expression strain, the expression strain is cultured and screened to obtain a cloned strain, and the cloned strain is shaken and packaged to obtain the hTERT-CDC6shRNA lentivirus; when the lentivirus disclosed by the invention is inoculated into a tumor body of a patient with craniopharyngeal tubuloma, the result shows that the lentivirus can effectively inhibit proliferation of craniopharyngeal tubuloma cells and can induce self apoptosis of the craniopharyngeal tubuloma cells.

The invention discloses an application of a Solyc11g071810 gene in regulating and controlling the size of tomato fruits, and is characterized in that the nucleotide sequence of cDNA (complementary deoxyribonucleic acid) of the Solyc11g071810 gene is as shown in SEQ ID NO. 1; according to the invention, a CRISPR / Cas9 gene editing technology is adopted, the Solyc11g071810 gene is edited, and an amino acid sequence coded by the Solyc11g071810 gene is changed, so that the function of the Solyc11g071810 gene disappears or changes, and the size of tomato fruits can be obviously reduced, therefore, the Solyc11g071810 gene can regulate and control the size of the tomato fruits, in addition, the invention also provides a CRISPR / Cas9 vector of the targeted Solyc11g071810 gene, and the expression of the Solyc11g071810 gene is improved by limiting a target site. And a better editing effect is achieved.

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 relates to the technical field of biological genes, in particular to clone of an oryza brachytonkinensis gene SBPL6 and application of the oryza brachytonkinensis gene SBPL6 to improvement of stigma traits of a rice male sterile line, in particular to application of the oryza brachytonkinensis gene SBPL6 to improvement of stigma traits of a sterile line, and improvement of the stigma traits of the sterile line is beneficial to improvement of outcrossing ability of the sterile line. The cDNA (complementary deoxyribonucleic acid) sequence of the gene SBPL6 is shown as SEQ ID NO.2. The wild brachyglossum gene SBPL6 can effectively increase the hair brush length and the stigma length of a rice stigma, so that the stigma exsertion rate is increased, and the outcrossing capability of a sterile line is favorably improved.

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.

The application of the primer for detecting the anti-disease gene of the Eriocheir sinensis in identifying the disease resistance of the parent of the Eriocheir sinensis comprises the following steps: (1) respectively extracting RNA (Ribonucleic Acid) of the Eriocheir sinensis, and carrying out reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid) of the Eriocheir sinensis; (2) designing a primer combination according to the sequence of the lysozyme gene of the eriocheir sinensis and the sequence of the C-type lectin 4 gene, performing PCR amplification on the cDNA sample of the eriocheir sinensis by utilizing the primer combination to obtain a target fragment, and detecting and confirming that the target fragment is amplified by utilizing agarose gel electrophoresis; (3) obtaining a cDNA sequence by a first-generation sequencing means, carrying out sequence comparison, identifying SNP sites in gene coding regions of the lysozyme gene and the C-type lectin 4 gene, and analyzing the two genes and a combination thereof; and comparing the differences of the disease resistance of the genotypes, and screening to obtain the genotype with the strongest disease resistance and the combined marker primer thereof.

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 belongs to the technical field of natural medicines, and particularly relates to a method for increasing the content of catharanthine in hairy roots of catharanthus roseus, which is characterized by comprising the following steps: extracting RNA (Ribonucleic Acid) of the catharanthus roseus, carrying out reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid), carrying out PCR (Polymerase Chain Reaction) amplification by taking the cDNA as a template to respectively obtain GS (Glutathione S) and CS (Citrus S), with the nucleotide sequence of the GS as shown in SEQ ID NO.9 and the nucleotide sequence of the CS as shown in SEQ ID NO.10; gS and CS are jointly cloned into an overexpression vector, and a double-gene expression vector is obtained; infecting the catharanthus roseus by using the double-gene expression vector; the infected catharanthus roseus is sequentially subjected to screening culture and subculture, an inducer is added in the subculture process, and the inducer is dexamethasone. Through the method provided by the invention, the content of catharanthine in the catharanthus roseus can be increased by 67% or above. The method for increasing the content of the vinblastine in the hairy roots of the catharanthus roseus is established, and a foundation is laid for large-scale production of the direct precursor vinblastine of an anti-cancer drug through the hairy roots.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-NKG2D nano antibody as well as a preparation method and application thereof. The method comprises the following steps: constructing and expressing NKG2D protein, immunizing alpaca to obtain immune alpaca PBMC (peripheral blood mononuclear cells), extracting RNA (ribonucleic acid) of the PBMC, carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), obtaining an antibody gene segment through PCR (polymerase chain reaction), and constructing the antibody gene segment on a phage expression vector; the method comprises the following steps: carrying out electrotransformation on a vector with an antibody gene segment to an SS320 electrotransformation competent state to obtain an NKG2D immune antibody library, carrying out two rounds of NKG2D protein panning and Clone ELISA detection to obtain an antibody combined with NKG2D protein, carrying out sequencing analysis, selecting a unique sequence, and carrying out high-throughput expression through a mammalian cell high-throughput expression system to obtain an antibody protein. And carrying out FACS detection by using an NKG2D high-expression cell line to finally obtain the antibody combined with the NKG2D. The method disclosed by the invention is simple and quick to operate; a mammalian cell expression system induces high-efficiency 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 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 discloses a method for detecting microbial genome RNA based on an mNGS technology. The invention provides a method for constructing an RNA sequencing library, which comprises the following steps: A1) fragmenting an RNA sample to obtain fragmented RNA; a2) carrying out reverse transcription and terminal repair reaction on the fragmented RNA in the following multi-enzyme reaction system to obtain double-chain cDNA (complementary deoxyribonucleic acid) with flat terminal and containing A basic group; and A3) sequentially carrying out joint connection and PCR amplification on the double-chain cDNA with the flat tail end and containing the A basic group to obtain the RNA sequencing library. According to the technical scheme, operation is easy, time consumption is short, reaction conditions and DNA library construction are the same, operation and reaction can be conducted at the same time, the initial amount of RNA is 100 pg, and daily requirements of microbiological detection are met.

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 targeted exosome loaded with siFKBP10 as well as a preparation method and application of the targeted exosome. The targeted exosome comprises a cell exosome with a membrane protein Lamp2b and a myofibroblast RDYH58 peptide which is combined with the membrane protein, and the cell exosome is modified by the siFKBP10; the cDNA (complementary deoxyribonucleic acid) sequence of the RDYH58 peptide is as shown in SEQ. ID. NO.1; 1, and the positive-sense strand of the siFKBP10 is as shown in SEQ ID NO. 2, and the antisense strand of the siFKBP10 is as shown in SEQ ID NO. 3. The targeting exosome can significantly reduce expression of FKBP10 in cells, inhibits excessive deposition of extracellular matrix, has a good inhibition effect on idiopathic pulmonary fibrosis, and provides a new treatment strategy for treatment of IPF.

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 belongs to the technical field of plant genetic engineering, and particularly relates to application of a tomato SlMYB78 gene to regulation and control of tomato resistance to pseudomonas syringae tomato pathogenic variants. The cDNA (complementary deoxyribonucleic acid) sequence of the SlMYB78 gene is as shown in SEQ ID NO. 1; the application approach is to weaken the resistance of PstDC3000 pathogenic bacteria by silencing the SlMYB78 gene. According to the invention, an SlMYB78 gene silenced transgenic tomato plant is obtained by using a gene silencing technology, and it is proved that after the SlMYB78 gene silenced strain is infected by PstDC3000 pathogenic bacteria, the accumulation of phenol amine metabolites and the synthesis of SA are reduced, and the disease resistance is significantly reduced.