35 results about "Genetic strain" patented technology

The invention belongs to the field of molecular biology, and discloses application of a segment of isolated nucleotide sequence in construction of zebra fish with reduced intramuscular spines. The nucleotide sequence is shown as SEQ ID NO.1, by using a conventional method in the field, the SEQ ID NO.1 is taken as a target gene to perform gene mutation operation, F0 with mutation is screened out, F1 generated by hybridization of F0 and a wild type is subjected to genotyping, and mutant F1 is screened out and selfed, so that the zebra fish strain with few intramuscular spines which is remarkablyreduced by more than 70% in the number of intramuscular spines and is stably inherited can be obtained. According to the application of the segment of isolated nucleotide sequence in construction ofzebra fish with reduced intramuscular spines, the segment of isolated nucleotide sequence is used to obtain a fish strain with reduced intramuscular spines for the first time, and the application of the segment of isolated nucleotide sequence in construction of zebra fish with reduced intramuscular spines has the advantages of remarkable character improvement effect, short breeding time, stable heredity, low cost and the like, has the basic value and the application value of scientific research, can be used for researching the development mechanism of the intramuscular spines of the fish, andprovides a method for obtaining a stable genetic strain with few intramuscular spines of various fish.

The invention belongs to the technical field of plant genetic engineering, and specifically designs a carrier for improving the regeneration efficiency of strawberry leaves and application of the carrier. An RNA interference fragments of a forest strawberry cytokinin receptor gene FveHK2 is screened from a forest strawberry, a plant inhibition expression vector GN2300-FveHK2 of the forest strawberry cytokinin receptor gene FveHK2 is constructed, then transfection is performed on transgenic engineering bacteria, the plant inhibition expression vector GN2300-FveHK2 is transferred into a diploidforest strawberry 'Hawaii 4' to obtain an RNAi expression interference strain line of the FveHK2 gene, under the condition of in-vitro leaf tissue culture, the germination time of the FveHK2 gene inhibition expression line is earlier than that of a wild type, and the budding rate of the FveHK2 gene inhibition expression line is significantly higher than that of the wild type, which indicates thatthe regeneration ability of the leaves of the forest strawberry transgenic line with the genetic transformation FveHK2 interference vector is significantly improved. The carrier for improving the regeneration efficiency of the strawberry leaves has important significance for the regeneration of strawberry leaves and the improvement of the transformation efficiency.

The invention discloses a method for breeding leaf mustard type rape varieties and materials by a rape double haploid inducing line. The method comprises the following steps: firstly, determining objective traits of the bred leaf mustard type rape varieties and materials; secondly, performing hybridization, convergent cross or backcross on at least two leaf mustard type rapes with the objective traits; thirdly, pollinating progenies obtained by hybridization, convergent cross or backcross by using the rape double haploid inducing line; fourthly, identifying the genetic stability of induced progenies, and realizing selfing breeding according to strains; fifthly, identifying the yield, resistance and quality traits of induced stable progeny strains; sixthly, determining the stable strains according to the yield, the quality traits and the resistance to form a conventional variety of leaf mustard type rape; seventhly, inducing stable progeny genetic strains to form a new leaf mustard type rape line and performing test cross on the new leaf mustard type rape line and a leaf mustard type rape sterile line to form a maintainer line or restorer line, or entering next round of breeding process of the varieties and materials. According to the method, the breeding speed and efficiency of a hybrid variety or a conventional variety of the leaf mustard type rape can be greatly improved, and labor force and material resources are reduced.

The present invention discloses a safe and environmentally friendly cichorium intybus chloroplast transformation system establishment method, which comprises: (1) establishing a high frequency regeneration system of a cichorium intybus leaf explant; (2) constructing a safe and environmentally friendly cichorium intybus chloroplast site-specific integration expression vector; and (3) adopting a gene gun bombardment method to transform the cichorium intybus explant, and carrying out three rounds of resistance screening to obtain the chloroplast transgene strain. According to the present invention, the novel safe selection marker gene PMI is adopted to replace the traditional antibiotic marker gene aadA to construct the chloroplast site-specific integration expression vector and establish the cichorium intybus chloroplast transformation system, wherein the system has advantages of high screening efficiency, high exogenous gene expression level, strong transgenic plant bio-safety and the like; and receptor types of chloroplast transformation are broadened, and technical platforms are built for genetic improvement on cichorium intybus, or application of cichorium intybus chloroplast as a bioreactor to produce animal oral vaccines, medical proteins and the like.

The invention discloses brassica napus L promoter pBnUnng0942890 and application thereof. By cloning a promoter of a gene pBnUnng0942890 in the brassica napus L, a plant expression vector of a reporter gene GUS regulated by the promoter is constructed; by using an agrobacterium-mediated inflorescence impregnating method to transform arabidopsis, GUS histochemical staining is carried out on screened positive transgenic strains; the result shows that the pBnUnng0942890 has the functions of driving preferential expression of downstream genes in roots, flowers and seeds, and not expressing or realizing low expression in other tissues and organs. The promoter has good application potential in the aspects of improving crop quality by rapeseed transgene, artificially creating germplasm resourcesand the like.

The invention discloses a selection method for a good genetic strain of sophora japonica, and belongs to the technical field of tree good strain selection. The method is capable of solving the problemin the prior art that the good genetic strain of the sophora japonica cannot be selected. The method comprises the following steps: after the primary selection and repeated selection, selecting superior trees, marking the superior trees, and breeding the superior trees through grafting and budding; and after the bred superior trees are in the full bearing period, performing the contrastive analysis to all tested clones, and selecting the superior tree clones which can keep the female parent good feature or achieve the local main cultural variety standard as the good strains. The method is capable of improving the variety rate of the sophora japonica, and optimizing the sophora japonica resources.

The invention belongs to the field of plant transgenosis and relates to a method for rapid screening of a single copy transgenic offspring homozygous family and a use thereof. A carrier is conversed through an agrobacterium-mediated genetic transformation method, a positive single copy transgenic plant is screened through southern, seeds of the T0 generation of the transgenic plant are collected and then are sowed in a seed bed, leaves in a trefoil stage are collected, total DNAs of a hand sample are extracted through a CTAB method, real-time PCR primers are designed according to an exogenous target gene hygromycin and an endogenous G3PDH gene, the hand sample DNA is subjected to real-time PCR through the primers of the exogenous target gene and the endogenous gene, amplification Ct values of the exogenous target gene and the endogenous gene are calculated and compared, a transgenic plant genotype is determined through the amplification Ct values, after the T1 generation of the plants produce fruits, a transgenic plant genotype is detected through germination, the above two transgenic plant genotypes are compared, and the two materials with the identical genotypes are used as transgenic breeding, hybridization transformation breeding or polygene pyramiding breeding materials.

The invention belongs to the technical field of biological genetic engineering and particularly discloses an oryza sativa histone methyltransferase as well as an encoding gene and an application thereof. The methyltransferase adopts one of the following amino acid residue sequences: (1) SEQ ID NO.1 in a sequence table; (2) substitution and / or deletion and / or addition of 1-50 amino acid residues performed on the amino acid residue sequence shown in the SEQ ID NO.1 in the sequence table, the methyltransferase is a protein adopting the sequence and having a regulation effect on growth and development of plants. The invention further discloses a recombinant expression vector, a transgenic cell line and engineering bacteria containing the gene as well as a primer pair for amplifying any fragment of the gene. Antisense transgenic plants of the encoding gene of the oryza sativa histone lysine methyltransferase grow short and small, and the flowering time is delayed. One excellent gene is provided for improvement of plant varieties and has higher actual application value and broad application prospect.

The invention relates to a GhMYB36 gene for verticillium wilt resistance of plants and application of the GhMYB36 gene, and belongs to the technical field of biology. The GhMYB36 gene is an R2R3 typeMYB protein gene obtained from upland cotton 3503 varieties and contains 2 DNA (deoxyribonucleic acid) binding domains. The GhMYB36 gene and the application have the advantages that the verticillium wilt resistance of GhMYB36 transgenic expression strains can be obviously improved, the incidence can be delayed, and the average incidence rate can be lowered to a great extent; the verticillium wiltresistance of transgenic strain lines can be obviously enhanced for defoliating type verticillium wilt after pathogenic bacteria are inoculated.

The invention relates to the field of plant molecular biology, in particular to a C. praecox CpUFO gene as well as coded protein and an application thereof. A cDNA sequence of the C. praecox CpUFO is obtained by gene cloning, the whole length of the cDNA sequence of the gene is 1753 bp, and the gene contains 1212 bp complete ORF (open reading frame) and codes 403 amino acids. After the gene is transformed into arabidopsis thaliana, phenotype observation finds that the bud emergence time, tillering time, first flower blooming time and first pod emergence time of a transgenic line are earlier than a WT plant; the rosette leaf number and total leaf number are fewer than the WT plant, and the blooming time is early. Flower organ observation finds that arabidopsis thaliana pistils genetically modified with the CpUFO gene at the same blooming stage are longer than wild type arabidopsis thaliana, which indicates that the C. praecox CpUFO gene can promote plants to bloom and has influence on flower organs of plants.

The invention discloses an artificial recombinant H5N8 influenza virus and a preparation method and application thereof. The artificial recombinant H5N8 influenza virus has a highly pathogenic avian influenza virus A / swan / Shanxi / 4-1 / 2020 (H5N8) surface antigen Hemagglutinin (HA) gene and a Neuraminidase (NA) gene, but an HA cleavage site has a typical low pathogenic avian influenza virus molecular characteristic (-RETR-), and has six internal genes of PB2, PB1, PA, NP, M and NS of an influenza virus chick embryo high-titer adaptive strain A / PR / 8 / 34(H1N1), the strain number is H5-Re14, and the preservation number is CCTCC NO: V202160. The artificial recombinant H5N8 influenza virus has good chick embryo adaptability, and the virus growth titer can be increased by 9 times or above compared with that of a wild virus; and the artificial recombinant H5N8 influenza virus is used as a vaccine strain for producing a vaccine, the production cost of the vaccine can be greatly reduced, and the quality of the vaccine is improved.

The invention discloses an artificial recombinant H7N9 influenza virus and a preparation method and application thereof. The artificial recombinant H7N9 influenza virus has a highly pathogenic avian influenza virus A / chicken / Yunnan / SD024 / 2021(H7N9) surface antigen Hemagglutinin (HA) gene and a Neuraminidase (NA) gene, but the amino acid sequence of the HA cleavage site is -PKGR-, the virus has six internal genes of PB2, PB1, PA, NP, M and NS of influenza virus A / PR / 8 / 34(H1N1), the strain number is H7-Re4, and the preservation number is CCTCC NO: V202161. The strain has good chick embryo adaptability, and the virus growth titer can be increased by 9 times or above compared with that of a wild virus; the strain is used as a vaccine strain to produce vaccines, the production cost of the vaccines can be greatly reduced, and the quality of the vaccines is improved.

The invention provides a novel array method for nucleic acid sequence detection with improved specificity which allows for detection of genetic variation, from simple SNPs (where the variation occurs at a fixed position and is of limited allelic number) to more complex sequence variation patterns (such as with multigene families or multiple genetic strains of an organism where the sequence variation between the individual members is neither fixed nor consistent). The array is comprised of short, synthetic oligonucleotide probes attached to a solid surface which are hybridized to single-stranded targets. Single stranded targets can be produced using a method that employs primers modified on the 5′ end to prohibit degradation by a 5′-exonuclease that is introduced to degrade the unprotected strand. The invention further provides for printing buffers / solutions for the immobilization of oligonucleotide probes to an array surface. The invention also provides hybridization and wash buffers and conditions to maximize hybridization specificity and signal intensity, and reduce hybridization times.

The invention provides a Trichoderma reesei strain with high cellulase-producing space mutagenesis, and belongs to the technical field of microbial mutation. In the present invention, the Trichoderma reesei Rw‑X1 strain is preferably equipped with "Shenzhou No. 9" for space mutagenesis, and the Trichoderma reesei (Trichoderma reesei) TG‑C521 strain capable of high cellulase production is cultivated after careful screening. On August 25, 2014, it was deposited in the China General Microorganism Culture Collection Management Center, and the preservation number is CGMCC No.9537. The present invention uses a special environment to change the genetic shape of the bacterial species, and optimizes positive variation and stable genetic strains. This bacterial strain is used to produce cellulase, and its filter paper enzyme activity is 25.83% higher than that of Trichoderma reesei Rw‑X1 strain ~34.49%, environmental protection and pollution-free, has broad industrial application prospects.

An isolated zebrafish genetic strain having a dystrophin mutant phenotype and fish models useful for screening or assaying agents having potential activity on muscular dystrophy or cardiomyopathy.

An isolated zebrafish genetic strain having a dystrophin mutant phenotype and fish models useful for screening or assaying agents having potential activity on muscular dystrophy or cardiomyopathy.

The present invention describes the use of a genetically improved Trichoderma reesei strain allowing to limit microbiological contaminations during an industrial process. The genetic strain improvement allows the latter to overexpress an extracellular protein having known antimicrobial properties and compatible with the secretory system of strains of fungi such as Trichoderma reesei. This modified strain can be used to produce the cellulolytic and / or hemicellulolytic enzymes used in a method of producing ethanol from cellulosic or lignocellulosic materials referred to as “second generation” materials.

The invention relates to a rice female genic male sterility gene engineering expression vector and application thereof, and belongs to the technical field of agricultural biology. In order to realize the breeding of seeds and seedlings of the rice female genic male sterile line, the invention provides a technical means of transgene transfer deletion to realize the breeding of a non-transgene homozygous female genic male sterile line. Specifically, after selfing and fruiting of a genetic engineering female genic male sterile line, namely a hybrid type transgenic T0 strain, a transgenic T1 segregation population is obtained, in the seedling stage of the transgenic T1 segregation population, transgenosis female fertility recovery plants are screened out by spraying bentazone herbicide, and survival T1 plants are non-transgenosis female genic male sterile (fst / fst) plants. The non-transgenic female genic male sterile line can meet the mechanical seed production operation of a hybrid rice seed production technical system of 'FS + MS (FM-line)', namely 'female sterile line + male sterile line (FM line method)'.

The invention discloses an artificially recombined H5N8 influenza virus, a preparation method and application thereof. An artificial recombinant H5N8 influenza virus of the present invention has highly pathogenic avian influenza virus A / swan / Shanxi / 4‑1 / 2020 (H5N8) surface antigen hemagglutinin (HA) and neuraminidase (NA) genes , but the HA cleavage site shows the molecular characteristics of typical low pathogenic avian influenza virus (‑RETR‑), and has the PB2, PB1, PA, There are 6 internal genes including NP, M and NS, the strain number is H5‑Re14, and the deposit number is CCTCC NO: V202160. It has good chicken embryo adaptability, and the virus growth titer can be increased by more than 9 times compared with wild virus; using this as a vaccine strain to produce vaccines can greatly reduce vaccine production costs and improve vaccine quality.

The invention discloses a type 3 duck hepatitis A virus mutant gene ISA-T1142A and its construction method. The 3403rd nucleotide G of the mutant gene genome is mutated into T, which can be used as a genetic marker for infectious clones and can be used to distinguish rescued strains With the parental strain and the wild strain; the 1142nd nucleotide is mutated from T to A, so that the 164th amino acid of the virus VPO protein is mutated from the tyrosine of the parental strain to asparagine. The mutant gene virus strain has the same antigenicity as the parent strain, and has higher proliferation efficiency and virus titer than the parent strain on duck embryos, and has good immunogenicity and genetic stability. The pathogenicity of the mutant gene virus strain to ducklings has been significantly reduced, which has laid an important foundation for the pathogenic mechanism of the virus and the development of vaccines. In conclusion, the type 3 duck hepatitis A virus mutant gene ISA‑T1142A virus strain can be used as a vaccine candidate or strain and can be used for the study of the attenuation mechanism of type 3 duck hepatitis A virus, with broad application prospects.

The invention discloses an artificially recombined H7N9 influenza virus, a preparation method and application thereof. The artificial recombinant H7N9 influenza virus of the present invention has highly pathogenic avian influenza virus A / chicken / Yunnan / SD024 / 2021 (H7N9) surface antigen hemagglutinin (HA) and neuraminidase (NA) gene, but HA cracks The amino acid sequence of the site is ‑PKGR‑, with 6 internal genes including PB2, PB1, PA, NP, M and NS of influenza virus A / PR / 8 / 34 (H1N1), the strain number is H7‑Re4, and the deposit number is CCTCC NO: V202161. The virus strain has good adaptability to chicken embryos, and the virus growth titer can be increased by more than 9 times compared with the wild virus; it can be used as a vaccine strain to produce vaccines, which can greatly reduce the production cost of vaccines and improve the quality of vaccines.

The invention discloses an antagonistic bacteria Z‑18 for preventing and treating Verticillium wilt of cotton and its application. Based on the relationship between plant endophytes and plant symbionts, a batch of endophyte strains are isolated from cotton, and a strain of genetic Stable characteristics, good biocontrol Bacillus ( Bacillus zhangzhouensis ) Z-18 CGMCC No.15006, when the screened endophytic strain Z-18 is used in the biological control of cotton Verticillium wilt, it can reduce the disease index of cotton Verticillium wilt, and the control effect is 59.5%. Cotton verticillium wilt, the main cotton disease, has a good control effect, and there is a significant difference (P<0.05) compared with the control, which becomes a new way to control cotton verticillium wilt, which is conducive to improving the yield and quality of cotton.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a carrier for delaying fragaria X ananassa Duch. leaf senescence and an application thereof. An RNA interference fragment of a fragaria X ananassa Duch. cytokinin receptor gene FveHK4 is screened from fragaria X ananassa Duch.; a plant inhibition expression vector GN2300-FveHK4 of the gene is constructed and is transferred into diploid fragaria X ananassa Duch. 'Hawaii 4', and an RNAi expression interference strain of the FveHK4 gene is obtained, wherein vegetative growth of the strain is notobviously different from vegetative growth of a wild type. Under a dark condition, compared with a wild type, senescence of the FveHK4-RNAi in-vitro leaves is obviously delayed, which shows that thesenescence speed of the leaves of the fragaria X ananassa Duch. transgenic line of the genetic transformation FveHK4 interference carrier is slowed down, and the functional period is prolonged. The carrier has important significance in delaying strawberry leaf senescence, increasing the photosynthetic production of leaves and further increasing the strawberry yield.

The invention relates to the technical field of plant disease control, in particular to the application of gene GhSINAs in the control of cotton verticillium wilt. The invention provides the application of gene GhSINAs in the prevention and treatment of cotton verticillium wilt, said gene GhSINAs comprises any one or more in gene GhSINA7, gene GhSINA8 and gene GhSINA9; The nucleic acid sequence of gene GhSINA7 is as SEQ ID NO:1 and / or shown in SEQ ID NO:2; the nucleic acid sequence of gene GhSINA8 is shown in SEQ ID NO:3 and / or SEQ ID NO:4; the nucleic acid sequence of gene GhSINA9 is shown in SEQ ID NO:5 and / or SEQ ID NO:6 shown. The present invention found in the research that among these three genes, each overexpressed transgenic line has improved resistance to Verticillium wilt, but the silencing of each gene has suppressed the defense ability to pathogen infection, indicating that GhSINA7, GhSINA8 and GhSINA9 play a positive regulatory role in cotton defense against Verticillium dahliae.

The invention belongs to the technical field of plant genetic engineering and specifically designs a carrier for improving regeneration efficiency of strawberry leaves and its application. The RNA interference fragment of forest strawberry cytokinin receptor gene FveHK2 was screened from forest strawberry, and the plant inhibitory expression vector GN2300‑FveHK2 of forest strawberry cytokinin receptor gene FveHK2 was constructed, and then transfected with genetically engineered bacteria, and transformed into two In the ploid forest strawberry 'Hawaii 4', the RNAi expression interference line of FveHK2 gene was obtained. Under the condition of isolated leaf tissue culture, the germination time of the FveHK2 gene suppressed expression line was earlier than that of the wild type, and the germination rate was significantly higher than that of the wild type. It indicated that the regenerative ability of the leaves of the transgenic line of strawberry transgenic line transformed with the FveHK2 interference vector was significantly improved. The invention has great significance to the regeneration of strawberry leaves and the improvement of transformation efficiency.

The invention discloses a method for breeding leaf mustard type rape varieties and materials by a rape double haploid inducing line. The method comprises the following steps: firstly, determining objective traits of the bred leaf mustard type rape varieties and materials; secondly, performing hybridization, convergent cross or backcross on at least two leaf mustard type rapes with the objective traits; thirdly, pollinating progenies obtained by hybridization, convergent cross or backcross by using the rape double haploid inducing line; fourthly, identifying the genetic stability of induced progenies, and realizing selfing breeding according to strains; fifthly, identifying the yield, resistance and quality traits of induced stable progeny strains; sixthly, determining the stable strains according to the yield, the quality traits and the resistance to form a conventional variety of leaf mustard type rape; seventhly, inducing stable progeny genetic strains to form a new leaf mustard type rape line and performing test cross on the new leaf mustard type rape line and a leaf mustard type rape sterile line to form a maintainer line or restorer line, or entering next round of breeding process of the varieties and materials. According to the method, the breeding speed and efficiency of a hybrid variety or a conventional variety of the leaf mustard type rape can be greatly improved, and labor force and material resources are reduced.