122 results about "Gene clone" patented technology

The invention discloses an SaRNA vaccine for echinococcosis as well as a preparation method and application of the SaRNA vaccine. The preparation method of the SaRNA vaccine comprises the following steps: carrying out codon optimization on a modified target antigen protein through a genetic engineering technology, then assembling the modified target antigen protein with a self-replicating protein sequence, 5 'UTR, 3' UTR and Poly (A) tail, carrying out gene synthesis, then cloning the synthesized gene into a plasmid, and carrying out purification to obtain the SaRNA vaccine. The preparation method comprises the following steps: constructing recombinant plasmids, sequentially carrying out plasmid linearization, in-vitro transcription and purification on the constructed recombinant plasmids to prepare SaRNA molecules, and finally wrapping the SaRNA molecules in lipid nanoparticles to form the SaRNA vaccine for the echinococcosis. Experiments prove that the SaRNA vaccine can activate humoral immunity and cellular immunity of mice at the same time, high-level EG95 specific antibodies and cytokines can be generated through low-dose immunity, and the SaRNA vaccine has wide application prospects in the aspect of preventing and / or treating the echinococcosis.

The invention discloses an echinococcosis mRNA (messenger Ribonucleic Acid) vaccine as well as a preparation method and application thereof. The preparation method of the mRNA vaccine for the echinococcosis comprises the following steps: carrying out codon optimization on a modified target antigen protein, then assembling the modified target antigen protein with 5 'UTR, 3' UTR and Poly (A) tail, carrying out gene synthesis, then cloning the synthesized gene into a pUC57 plasmid, and sequentially carrying out plasmid linearization, in-vitro transcription and purification on the constructed recombinant plasmid to prepare an mRNA molecule, thereby obtaining the mRNA vaccine for the echinococcosis. Finally, mRNA molecules are wrapped in lipid nanoparticles through a microfluidic method to form the mRNA vaccine for the echinococcosis, and immune effect evaluation is carried out on the mRNA vaccine for the echinococcosis. Experiments prove that the prepared mRNA vaccine for the echinococcosis can activate humoral immunity and cellular immunity of mice at the same time, can provide an effective protection effect for the mice attacking insects, and has a wide application prospect in the aspect of preventing and / or treating the echinococcosis.

The invention provides the cassava low-temperature-resistant MeSR45-2 gene cloned from cassava for the first time, the expression of the MeSR45-2 gene in cassava is inhibited under low-temperature stress, and research shows that the gene can significantly improve the low-temperature resistance of cassava, reduce the malondialdehyde content, proline cumulant and the like in cassava, reduce leaf damage under low-temperature treatment and improve the yield of cassava. Terminal buds under low-temperature treatment are not affected and can normally grow, and expression of various protein genes is regulated and controlled. The invention provides a new candidate gene for research on improvement of plant low temperature resistance and the like, and also provides a new gene resource for cultivation of a new variety of low temperature resistant cassava and construction of a new variety of transgenic cassava.

The invention discloses a preparation method and application of a cat main allergen FEL-1 nano antibody FEL-1-Nb, and relates to the technical field of antibody preparation, and the preparation method comprises the following steps: preparing an immunogen; animal immunization and library construction; constructing and panning a phage display library; screening and identifying positive clone; and expression and purification of the nano antibody: cloning the positively cloned VHH gene to an expression vector such as a pET series, transforming an expression strain such as escherichia coli BL21 (DE3), carrying out IPTG induced expression, and purifying through an inclusion body purification method to obtain a high-purity nano antibody protein. The nano antibody FEL-1-Nb provided by the invention can be specifically bound with a cat main allergen FEL-1 protein, the unique amino acid sequence of the nano antibody FEL-1-Nb is shown as SEQIDNO: 1, high-affinity binding with the FEL-1 protein is ensured, the positive clone binding rate is high through phage display panning and ELISA verification, the phage recovery rate after the third round of panning is increased by more than 1000 times compared with the first round of panning, and the positive clone binding rate is higher than that of the first round of panning. Therefore, the nano antibody has excellent targeted recognition capability.

The invention belongs to the technical field of plant genetic engineering and molecular breeding, and particularly relates to application of a soybean GmUGT72E1 gene in regulation and control of soybean plant height and enhancement of resistance of soybeans to drought and alkali stress. Plant height is a key agronomic character influencing lodging resistance and photosynthetic efficiency of soybeans, drought and alkali stress are core abiotic stress factors restricting yield and quality of the soybeans, and the three factors jointly lead to remarkable global soybean yield reduction loss. Gene cloning, vector construction and transgenic function verification prove that overexpression of the GmUGT72E1 gene can reduce the plant height of soybeans by 15%-25%, meanwhile, the tolerance of the soybeans to drought and alkali stress is remarkably improved, the relative water content of leaves under the drought stress is increased by 25%-30% compared with that of wild type leaves, and chlorophyll under the alkali stress is increased by 40%-50% compared with that of the wild type leaves. According to the application disclosed by the invention, the dual regulation and control functions of the GmUGT72E1 gene are defined, key gene resources and technical support are provided for soybean dwarf stress-resistant collaborative improvement, dwarf stress-resistant soybean varieties can be cultivated by means of transgenosis, gene editing and the like, and the application has important agricultural application value.

The invention discloses a method for synthesizing alpha-arbutin through biotransformation, and belongs to the technical field of biochemistry. A gene cloning technology is utilized, sucrose phosphorylase Spase in leuconostoc mesenteroides and sucrose phosphorylase gtfa in bifidobacterium adolescentis are screened and found through a Deep Molecules and DLKcat algorithm on a Uniprot database, novopro is utilized for codon optimization, then the sucrose phosphorylase Spase and the sucrose phosphorylase gtfa are expressed on a plasmid vector PET-28a and are converted into an escherichia coli BL21 (DE3) strain, and the sucrose phosphorylase Spase and the sucrose phosphorylase gtfa in the bifidobacterium adolescentis are obtained. An engineering strain for heterologous expression of the gene is constructed by taking the gene as a chassis cell. According to the process, sucrose and hydroquinone are taken as co-substrates, and bioconversion is carried out under the action of sucrose phosphorylase from different sources to obtain alpha-arbutin.

The application discloses a transcription factor for regulating protein content of rice grains GPC6 and application, the amino acid sequence of the transcription factor is shown as SEQ ID NO. 1, the transcription factor comprises a genomic region and a promoter, the nucleotide sequence of the genomic region is shown as SEQ ID NO. 2, the nucleotide sequence of the promoter is shown as SEQ ID NO. 3, the coding sequence (CDS) is shown as SEQ ID NO. 4, and all alleles thereof are contained. The application identifies a repeatedly detectable locus on chromosome 6 by performing whole genome association analysis on gluten content and total protein content GPC6 . Gene cloning is carried out on the locus, so that the genetic resources for improving rice quality are enriched, and help is provided for improving rice quality, in particular, protein content.

This invention discloses a SUMO-C7H 15 This invention relates to the field of biotechnology, specifically to NO2S probes, their synthesis methods, and applications. The molecular formula of the probe described in this invention is SUMO-C7H. 15 NO2S, wherein the SUMO is human SUMO-1; the nucleotide sequence of the human SUMO-1 is shown in SEQ ID NO: 1, and the amino acid sequence is shown in SEQ ID NO: 2. This invention obtains the polypeptide sequence encoding the human SUMO-1 protein through gene cloning technology, and further prepares a structurally stable SUMO-C7H protein using chemical synthesis methods. 15 NO2S, this probe has a stable molecular structure and is easy to synthesize and detect; SUMO-C7H 15 The NO2S fluorescent probe possesses high specificity and good detection sensitivity, providing an innovative research tool for exploring tumorigenesis mechanisms and assessing SUMOylation modification levels.

The application discloses a preparation method of a recombinant ternary fusion protein dressing with epidermal cell activation function. The dressing is a fusion protein designed based on the research on the functions and properties of COL3A1 and IFN-kappa proteins by international peers and the laboratory. The specific method is that the fusion gene fragment of the covalent connection of COL3A1 and IFN-kappa with GGSGG as a linker is obtained by using overlap extension PCR technology, and a 6×His label is added. The target gene is cloned into a prokaryotic expression vector pET30 (modified in the laboratory) to obtain the fusion protein COL3A1-IFN-kappa with an MBP label. After expansion culture, the MBP-IFN-kappa-COL3A1 with a purity of more than 90% can be obtained through Ni-NTA column affinity chromatography. We verified the resistance of the fusion protein to salmonella in THP-1 cells, and verified the effect of the fusion protein on promoting wound healing and resisting salmonella infection on mice. Finally, it is proved that the MBP-IFN-kappa-COL3A1 has good uniformity, is easy to purify, has high activity, has no side effects, and is very suitable for being used as an epidermal cell activation dressing.

The present invention relates to a multi-assembly platform capable of manufacturing a plurality of fragmented level 1 transcription units in a single reaction using connectors, and then assembling the plurality of level 1 transcription units at once by using yeast homologous recombination technology. The present invention has an effect of significantly improving assembly efficiency while simplifying procedures compared to existing methods. Therefore, the present invention can be effectively used in the synthetic biology and biofoundry fields requiring diverse and high-throughput gene cloning products.

The invention provides a GMRM (Genome-based Mutant RNA Mapping) method taking a genome as a reference, in particular, the invention provides a method for cloning candidate functional genes of wheat stripe rust resistant plants and an analysis process, and the GMRM method disclosed by the invention can be used for simultaneously cloning two genes with complementary disease-resistant functions, namely Yr6NLR1 and Yr6NLR2. In an example of Yr6, the two disease-resistant genes are closely linked genetically, but the cloning process of the genes does not involve any genetic recombination. Therefore, theoretically, the method provided by the invention can be used for simultaneously cloning a plurality of functional complementary genes (namely genes participating in the same pathway) on the premise that the target gene can generate effective mutation through EMS mutagenesis.

The invention discloses a preparation method and application of carassius auratus rahn aquatic OmpA gene cloning and prokaryotic expression as well as a polyclonal antibody. The preparation method comprises the following steps: culture and DNA amplification of rahn aquatic, design of an OmpA primer, gene cloning and prokaryotic expression, back multi-point subcutaneous injection of an immune rabbit, heart blood sampling and preparation of the polyclonal antibody. The invention further provides OmpA protein prokaryotic expression and protein purification, preparation of OmpA polyclonal antiserum and application of fish bodies. According to the invention, a rahnella aquatilis OmpA gene is cloned, a specific Hind III and EcoR1 double-enzyme digestion primer sequence OmpA-1F / OmpA-1R is designed, a prokaryotic expression vector pET32a-OmpA is constructed, high-immunogenicity OmpA protein is obtained through purification, OmpA specific multi-antibody serum is prepared, and the OmpA specific multi-antibody serum can be used for preparing the multi-antibody serum. The OmpA polyclonal antibody can be used for detecting and positioning the expression application of rahnella aquatica OmpA protein in crucian carp bodies through bidirectional agar diffusion, an immunoblotting method and an immunohistochemical method, and provides a reference basis for research and development and application of rahnella aquatica subunit vaccines and a molecular diagnosis technology as well as immune prevention and control of infectious diseases of the rahnella aquatica subunit vaccines and the molecular diagnosis technology.

The present application belongs to the technical field of plant genetic engineering, and particularly relates to a Phyllostachys edulis PeMASR1 gene, biological material and application thereof. The PeMASR1 gene provided by the present application is a new gene discovered in the study of cold tolerance of Phyllostachys edulis leaves, the nucleotide sequence of which is shown as SEQ ID NO:1, and the amino acid sequence of the encoded protein is shown as SEQ ID NO:2. Through gene cloning, expression and comparative analysis, the phylogenetic position of the Phyllostachys edulis PeMASR1 gene and the important role thereof in the research of plant response to abiotic stress mechanism and photosynthesis are revealed. Overexpression in Columbia-type Arabidopsis plants can make the rosette leaves of the plants early senesce; and the plants show a more stress-tolerant phenotype than wild-type Arabidopsis in response to cold stress, drought stress and salt stress and other abiotic stresses.

The invention belongs to the technical field of detection devices, and discloses a detection device and a rapid detection method for enteromorpha microscopic propagules. According to the detection device, a connector and a sample feeding pipe and a connecting pipe at the bottom and the top of a filter are limited through a first protection assembly, a second protection assembly and a limiting assembly respectively, and meanwhile, protection blocks are used for protecting the outer sides of the first protection assembly, the second protection assembly and the filter, so that the filter is prevented from being easily loosened due to vibration during operation of the filter; the rapid detection method comprises the following steps: 1, filtering a collected water sample through a filter membrane in a filter instrument, and extracting high-purity DNA (deoxyribonucleic acid); 2, cloning and purifying the enteromorpha specific gene, determining the DNA concentration of the target gene, and establishing a standard curve; and 3, detecting the copy number of the enteromorpha microcosmic propagule in the water body through qPCR, and calculating the cell abundance according to the copy number of the target gene in a single cell to realize rapid identification and quantitative detection.

The application provides application of a GmERF5 gene in drought resistance and salt stress resistance, and belongs to the technical field of genetic engineering.The GmERF5 gene is cloned into a pCAMBIA3301 vector to obtain a recombinant vector; the obtained recombinant vector is transformed into Agrobacterium rhizogenes, and a transgenic complex plant is obtained by using the obtained Agrobacterium rhizogenes to infect seeds, so that overexpression of the GmERF5 gene is realized.The overexpression of the GmERF5 gene can improve the antioxidant enzyme activity of the transgenic soybean plant, and the drought and salt stress resistance of the soybean is enhanced.The soybean GmERF5 of the application provides a new regulation gene resource for improving the drought resistance and salt tolerance of soybean, can be used for cultivation and improvement of stress-resistant soybean materials, lays a theoretical foundation for drought resistance and salt molecular mechanism, and simultaneously provides a theoretical basis and gene resource for soybean drought resistance and salt molecular breeding.

This invention relates to the field of bioengineering technology, and more particularly to a gene for identifying salt tolerance in upland cotton, GhNRX16, and its molecular marker. The CDS sequence of the upland cotton gene GhNRX16 is shown in SEQ ID NO.1 of the sequence listing. This invention uses RNA-seq technology to screen and identify this salt tolerance-related gene in the transcriptome data of upland cotton under salt stress treatment. Furthermore, the GhNRX16 sequence was obtained through gene cloning, and a transient silencing vector based on VIGS (virus-induced gene silencing) technology was successfully constructed. Experiments using VIGS-mediated gene silencing combined with salt stress treatment showed that silencing the GhNRX16 gene significantly reduced the plant's tolerance to salt stress, demonstrating that normal expression of this gene directly contributes to ensuring the plant's salt stress resistance. These results confirm the crucial role of the GhNRX16 gene in regulating cotton salt tolerance, providing important gene resources and theoretical basis for molecular breeding of salt-tolerant cotton.

This invention relates to the field of crop genetics and breeding, and discloses the sweet sorghum disease resistance gene CPK4, its encoded protein, and its applications. The nucleotide sequence of the sweet sorghum disease resistance gene CPK4 is shown in SEQ ID NO.1, or has more than 90% homology with the sequence shown in SEQ ID NO.1 and encodes a protein with a nucleotide sequence that promotes the release of reactive oxygen species (ROS) in plants. By utilizing the CPK4 gene cloned from sweet sorghum, and through the specific interaction between its encoded protein and the NADPH oxidase SbRbohD protein, the ROS signaling pathway induced by pathogens is specifically activated, strengthening the defense response of sweet sorghum against anthracnose. This achieves a targeted enhancement of sweet sorghum's resistance to anthracnose. Furthermore, this gene is specifically induced by the molecular patterns of anthracnose and pathogens, and its disease resistance mechanism differs from existing NLR-mediated immune pathways, enriching the types of sweet sorghum disease resistance gene resources.

This invention discloses a Panax notoginseng ethylene response factor gene. PnERF120 Its nucleotide sequence is shown in SEQ ID NO:1, and the encoded amino acid sequence is shown in SEQ ID NO:2. PnERF120 The gene originates from Panax notoginseng, and the encoded protein sequence contains a typical conserved domain of the ERF transcription factor. This invention demonstrates, through gene cloning, bioinformatics analysis, expression pattern analysis, subcellular localization, overexpression, and other techniques and methods, that... PnERF120 This invention can be used to improve the resistance of Panax notoginseng to Panax notoginseng A virus, and also to improve the resistance of tobacco to tobacco mosaic virus. This invention provides new functional genes and application pathways for antiviral breeding and disease-resistant materials of Panax notoginseng and other plants.

The present application relates to the technical field of biological genetic engineering, and particularly relates to a kind of identification of plant salt tolerance land cotton gene GhNRX71 And molecular marker, the CDS sequence of the land cotton gene GhNRX71 As shown in SEQ ID NO.1 In the sequence table.The present application is screened and identified in the transcriptome data of land cotton under salt stress treatment by transcriptome sequencing (RNA-seq) technology.The sequence of GhNRX71 Is obtained by further gene cloning means, and the transient silencing vector based on VIGS (virus-induced gene silencing) technology is successfully constructed.The results show that the tolerance of plant to salt stress significantly reduces after silencing GhNRX71 Gene by using VIGS-mediated gene silencing combined with salt stress treatment experiment, which proves that the normal expression of the gene has a direct effect on guaranteeing the salt stress resistance of plant.The above results confirm that GhNRX71 Gene plays a key role in regulating the salt tolerance of cotton, and provides important gene resources and theoretical basis for cotton salt-tolerant molecular breeding.

The invention discloses an exocytosis protein subcellular localization method for mannitol-induced plasm-wall separation, and relates to the technical field of cytobiology, the method comprises the following specific steps: constructing a fusion expression vector: connecting a plant target exocytosis protein gene with a fluorescent protein gene through a gene cloning technology to obtain a fusion expression vector; inserting into a plant expression vector to obtain a recombinant expression vector containing the target extracellular secretory protein-fluorescent protein fusion gene; performing bacterial colony PCR identification and sequencing verification on the recombinant expression vector; by adopting an agrobacterium tumefaciens-mediated plant leaf instantaneous transformation technology, the fusion gene is rapidly expressed in plant cells, subsequent experiments can be carried out after culture is carried out for 2-3 days, the experiment period is greatly shortened, meanwhile, leaf cells are induced to be subjected to plasmon-wall separation through a gradient mannitol solution, positive cell screening and identification are not needed, and the application prospect is wide. And positioning can be realized by directly observing fluorescence signal distribution by using a laser confocal microscope.

The invention discloses recombinant lactobacillus casei for expressing sialyltransferase ST6GALNAC1, and belongs to the technical field of genetic engineering and microorganisms. The recombinant bacterium is obtained by cloning an ST6GALNAC1 gene to a pPG612 vector and electrically transforming the ST6GALNAC1 gene into lactobacillus casei ATCC393. Experiments show that the engineering bacterium can significantly enhance glycosylation modification of intestinal mucoprotein MUC2, improve expression of tight junction proteins (ZO-1, Occludin and Claudin-1), inhibit activation of a p38 / MAPK inflammation pathway and regulate an intestinal flora structure, so that intestinal injury and systemic inflammatory response caused by salmonella are effectively relieved. The invention provides a strain basis and technical support for developing a novel oral microecological preparation.

The application discloses a CMV virus-like particle for producing a VLP recombinant vaccine and a preparation method thereof, the CMV virus-like particle is obtained by cloning a C-terminal G4SLPETG modified plant virus cucumber mosaic virus CMV capsid gene into a prokaryotic expression vector to obtain a recombinant expression vector, the recombinant expression vector is transfected into E. coli BL21 (DE3), and expression is obtained through the recombinant E. coli BL21 (DE3); and the amino acid sequence of the C-terminal G4SLPETG modified CMV is SEQ ID NO. 1. Test proves that the recombinant strain constructed by the application is stable to expression of an exogenous protein. The recombinant protein expressed by the application is used for preparing a virus-like particle for antigen coupling, the coupling efficiency is high, the antigen purity is high, the safety is good, the virus-like particle is not pathogenic to mice and other animals, and the safety evaluation is easy.

This invention discloses a ribonuclease derived from tartary buckwheat. FtRNH Genes, their expression vectors, and applications. This invention isolates ribonuclease from tartary buckwheat through gene cloning. FtRNH The gene, whose CDS sequence is shown in SEQ ID No. 1, and whose encoded protein amino acid sequence is shown in SEQ ID No. 2, is described in this invention. FtRNH Transgenic lines were obtained by genetic transformation into cultivated tartary buckwheat (ZK3) and subjected to ultraviolet stress treatment. Compared with wild-type materials, the transgenic lines showed increased expression. FtRNH The positive lines of tartary buckwheat exhibited stronger tolerance to ultraviolet radiation and less accumulation of UV-B-induced R-loops and cyclobutanepyrimidine dimers. This invention has significant application prospects in improving the tolerance of cultivated tartary buckwheat to ultraviolet stress or in breeding tartary buckwheat varieties resistant to ultraviolet stress.

The invention discloses a CD147 nano antibody D2 as well as a preparation method and application thereof, and belongs to the technical field of molecular biology and immunology. The preparation method comprises the following steps: screening from an alpaca nano antibody phage library by utilizing a phage display technology, and sequencing to obtain 9 CD147 nano antibodies; 3 nano antibody genes are selected and cloned into pCold II, and prokaryotic expression recombinant plasmids are successfully constructed; the method comprises the following steps: carrying out inducible expression for 20 hours under the conditions that the final concentration of IPTG (isopropyl-beta-d-thiogalactoside) is 0.2 mmoL / L, the temperature is 16 DEG C and the speed is 100 rpm / min, carrying out nickel column purification, and then carrying out Coomassie brilliant blue dyeing and Western blot identification to determine and obtain the CD147 nano antibody which is named as C36, D2 and H4; the binding activity of the three nano antibodies and the CD147 protein is detected through indirect ELISA (enzyme-linked immuno sorbent assay). The result shows that the CD147 antigen and the three nano antibodies have binding activity and are dose-dependent. Under the same conditions, when the incubation concentration of the nano-antibody is as low as 1.25 micrograms / mL, the D2 binding effect is the best, the amino acid sequence of the nano-antibody is shown as SEQ ID NO.8, and the nucleotide sequence of the nano-antibody is shown as SEQ ID NO.10. The invention provides a more accurate method for detecting the CD147.

The invention discloses a CD147 nano antibody G45 as well as a preparation method and application thereof, and belongs to the technical field of molecular biology and immunology. A phage display technology is used for screening from a natural alpaca nano antibody phage library, finally three nano antibody genes are selected and cloned into pCold II, and prokaryotic expression recombinant plasmids are successfully constructed; the method comprises the following steps: carrying out inducible expression for 20 hours under the conditions that the final concentration of IPTG (isopropyl-beta-d-thiogalactoside) is 0.2 mmoL / L, the temperature is 16 DEG C and the speed is 100 rpm / min, carrying out nickel column purification, and then carrying out Coomassie brilliant blue dyeing and Western blot identification, so as to determine and obtain the CD147 nano antibody which is named as A8, B8 and G45; the binding activity of the three nano antibodies and the CD147 protein is detected through indirect ELISA (enzyme-linked immuno sorbent assay). The result shows that the CD147 antigen and the three nano antibodies have binding activity and are dose-dependent. Under the same conditions, when the incubation concentration of the nano-antibody is as low as about 0.625 g / mL, the G45 binding effect is good, the amino acid sequence of the nano-antibody is shown as SEQ ID NO.7, and the nucleotide sequence of the nano-antibody is shown as SEQ ID NO.10. The invention provides a more accurate method for high-sensitivity detection of CD147.

The invention discloses a quaternary ammonium salt PCR (Polymerase Chain Reaction) hot start reagent as well as a preparation method and application thereof, the chemical name of the PCR hot start reagent is 5 ', 5' '-bis (4-bromophenyl)-2', 2 '', 4 ', 4' ', 6', 6 ''-hexamethyl-1, 1 ': 3', 1 ': 3', 1 '''quaternary ammonium salt, and the PCR hot start reagent is a compound formed by removing two bromine substituents in a parent nucleus molecule with the CAS number of 1093856-24-6 of quaterphenyl quaternary ammonium salt. The PCR hot start reagent is suitable for gene cloning, clinical diagnosis, gene detection and other scenes with high specificity requirements, can effectively inhibit non-specific amplification at low temperature, improves the specificity of PCR reaction and product purity, and has popularization and application values.

The invention discloses a rape cold-resistant gene and application thereof, and belongs to the technical field of gene engineering. Four rape specific lone genes (BnaOGs) and 2859 brassica specific genes (BSGs) are identified through comparative genomics and multi-database artificial verification by combining UniProt-KB, Nrdb and the like, and the characteristics of short sequence, high GC content, no transcription factor and the like are found. Aiming at a BSG gene BnaA03G0334200ZS, a CDS sequence of the BSG gene BnaA03G0334200ZS is cloned, a pEGOEP35S-H overexpression vector containing a 35S strong promoter and a hygromycin selection marker is constructed, and a positive plant is obtained through agrobacterium-mediated transformation of brassica napus Westar. Functional verification shows that overexpression of the gene remarkably enhances the cold resistance of the rape, plant damage is slight at low temperature, the relative conductivity and MDA content are reduced, and the activities of SOD, POD and CAT are improved. The invention provides a new target and technical support for cultivation of cold-resistant rape varieties, and has a wide application prospect.

The invention discloses a gene BnA09GDSL for regulating and controlling male sterility of brassica napus and application of the gene BnA09GDSL. According to the invention, a novel gene BnA09GDSL for regulating and controlling the male sterility of the brassica napus is identified, a PC3301-BnA09GDSL overexpression vector and a CRISPR-BnA09GDSL gene editing vector are constructed, the gene BnA09GDSL is overexpressed in a male sterility line Bngms1, and the plant fertility is recovered; meanwhile, a CRISPR-BnA09GDSL transgenic plant is obtained, and the transgenic plant has a male sterility phenotype. The BnA09GDSL gene obtained by cloning can regulate and control the fertility of the brassica napus, so that cross breeding is facilitated, and the BnA09GDSL gene has important application value in the field of male sterility breeding of the brassica napus.

The invention provides construction and application of an intermolecular structural domain tension probe, the probe comprises a first structure, a second structure and a fluorescent protein pair eCFP and eYFP, the first structure and the second structure are respectively connected to two ends of the fluorescent protein pair, and the eCFP and the eYFP are connected through a short peptide; the first structure is a structural domain or protein capable of being combined with a cytoskeleton connecting protein, and the second structure is a structural domain or protein capable of being combined with a cytoskeleton; the cytoskeleton connecting protein is selected from talin, SYNE4, SYNE3, SYNE2, Desmoplakin and the like; the cytoskeleton is a microfilament, a microtubule or an intermediate filament. The intermolecular structural domain tension probe can be used for detecting intracellular mechanical signals, the limitation of a traditional tension probe is overcome, the adverse effect of function enhancement caused by gene cloning is avoided, a new opinion is provided for a mechanical transduction molecular mechanism, and the application has the advantages that the application range is wide, and the application prospect is wide. And a more accurate experimental tool is provided for the research of cell mechanics biology, and the effect of a mechanical signal in cell behavior regulation and control is disclosed.

The invention provides an anti-mycoplasma hyopneumoniae nano antibody and application thereof, and belongs to the technical field of biology. The amino acid sequence of the nano antibody is as shown in SEQ ID NO: 2. The invention also provides a coding gene of the nano antibody, a recombinant vector containing the coding gene, a recombinant strain and a recombinant cell. The preparation method of the nano-antibody comprises the following steps: cloning a coding gene of the nano-antibody into a vector, and then converting into a competent cell to obtain a recombinant strain for expressing the nano-antibody; after IPTG induced expression is adopted, the nano antibody is obtained. According to the present invention, the mycoplasma hyopneumoniae-oriented nano-antibody with the metabolic inhibition effect is firstly found, the metabolic inhibition valence is 105, the mycoplasma hyopneumoniae-oriented nano-antibody is used for preventing and / or treating mycoplasma hyopneumoniae infection, the effect is significant, and the use of antibiotics is reduced. The preparation method of the anti-mycoplasma hyopneumoniae nano antibody is simple, the cost is low, the yield reaches 70 mg / L, and the anti-mycoplasma hyopneumoniae nano antibody is suitable for large-scale production.