20 results about "Genetic Technique" patented technology

The application belongs to the technical field of aquatic genetics, and particularly relates to application of a group of SNP sites in detection of anti-NNV traits of Hippocampus kuda. The application provides a group of molecular marker combinations containing 680 core SNP sites, and sequences of the SNP sites are shown as SEQ ID NO. 1 to SEQ ID NO. 680. The SNP sites are obtained by whole genome resequencing of a reference population of H. kuda, and are screened by using MLE-rank algorithm to be significantly associated with anti-NNV traits. Based on the SNP combinations, the application further provides corresponding liquid phase chips, genomic selection breeding methods and kits. The technology can realize early and accurate evaluation of anti-NNV traits of H. kuda, significantly improve breeding efficiency, reduce detection cost, and provide an effective molecular breeding tool for breeding of H. kuda disease-resistant fine varieties.

The invention discloses a foot-and-mouth disease virus recombinant strain with a heat-resistant characteristic as well as a preparation method and application of the foot-and-mouth disease virus recombinant strain. The recombinant strain is obtained by rescuing foot-and-mouth disease virus infectious cDNA containing mutation sites through a reverse genetic technology, and the foot-and-mouth disease virus infectious cDNA containing mutation sites is obtained by mutating 4863rd nucleotide A into C through site-specific mutagenesis on the basis of a sequence shown in SEQ ID NO.1. The invention further discloses a preparation method of the foot-and-mouth disease virus infectious cDNA containing mutation sites. Experiments prove that compared with an original strain, the recombinant strain has higher heat resistance, the capsid complete splitting temperature of the recombinant strain is higher than that of the original strain, and after heat treatment, the proportion of complete virus particles of the recombinant strain is obviously higher than that of the original strain. Moreover, compared with an original strain, the recombinant strain shows better immunogenicity in a mouse body after being subjected to heat treatment at 37 DEG C. Therefore, the recombinant strain has an important application prospect in vaccine preparation, and a new technical means is provided for improving the heat stability of the vaccine and optimizing the foot-and-mouth disease prevention and control technology.

The invention relates to the technical field of poultry viruses, and discloses a pigeon I-type paramyxovirus attenuated strain aQY as well as a construction method and application thereof. According to the invention, a PPMV-1 QY strain genome is divided into five fragments A-E in full length by selecting a proper enzyme cutting site, a genetic marker is constructed through synonymous mutation based on a reverse genetic technology, the enzyme cutting site is added, the fragment A, the fragment B and the fragment E are properly modified, a full-length plasmid pOK-aQY is prepared, and then the full-length plasmid pOK-aQY and auxiliary plasmids pCI-NP, pCI-P and pCI-L are co-transfected to a BSR-T7 / 5 cell, so that the PPMV-1 QY strain is obtained. And after culture, taking cell supernatant, and inoculating the cell supernatant to a chick embryo to obtain the low virulent strain aQY of the pigeon type I paramyxovirus. The pigeon I-type paramyxovirus attenuated strain aQY strain has good stability, the MDT of the strain is 108 h, and the ICPI of the strain is 0. The method is suitable for research of PPMV-1 genome characteristics and preparation of pigeon Newcastle disease vaccines, and has important social benefits and outstanding application potential.

The application belongs to the technical field of biology, and particularly relates to a 3D protein mutant O foot-and-mouth disease recombinant virus strain and construction and application thereof. The application mutates methionine at the 403th position of the 3D protein of the O foot-and-mouth disease virus into alanine, and successfully constructs and rescues a 3D protein mutant O foot-and-mouth disease recombinant virus strain rFMDV-M403A by using reverse genetic technology. The amino acid of the recombinant virus strain can be stably mutated in the process of subculture, and the growth speed of the mutant strain is slower than that of the wild strain, and the virus titer is significantly reduced. In individual experiments, the infection of the mutant strain to suckling mice shows that the mutant strain can improve the survival rate of the suckling mice, and significantly reduce the viral load in the liver and the tissue lesions. The application provides an attenuated foot-and-mouth disease recombinant virus strain, which has important significance for in-depth understanding of the pathogenic mechanism and prevention and control strategy of the foot-and-mouth disease virus, and provides an important theoretical basis for further development of the foot-and-mouth disease genetic engineering vaccine.

The application provides a construction method of a recombinant canine distemper virus strain expressing mink parvovirus VP2 protein and relates to the technical field of genetic engineering. The construction method uses a mink canine distemper virus attenuated vaccine CDV3 strain as a basis, clones mink parvovirus VP2 genes into a canine distemper virus genome cDNA, and obtains a recombinant canine distemper virus rCDV3-mVP2 strain capable of simultaneously expressing CDV proteins and MEV VP2 proteins through virus reverse genetic technology, so as to provide a candidate vaccine strain for the research and development of a mink canine distemper and parvovirus enteritis double recombinant live vaccine.

The invention provides a closely linked SSR marker and application thereof in barley helminthosporium leaf spot resistant QTL positioning, and relates to the technical field of molecular marker assisted breeding. The two QTL sites closely linked with the helminthosporium leaf spot resistance character are successfully positioned through the SSR marker technology, the accuracy of the SSR marker is further verified, the positioning work of the barley helminthosporium leaf spot resistance QTL is successfully completed within a short time by utilizing the SSR marker technology, timely and effective support is provided for disease-resistant breeding, and the method has the advantages of being high in practicability and high in practicability. According to the calculated QTL effect, the position of the QTL on the genetic map is determined, fine positioning of the QTL can be achieved, cross breeding is conducted through the screened helminthosporium leaf spot resistant genotype, a new helminthosporium leaf spot resistant barley variety can be more quickly cultivated in combination with a traditional breeding method, and the strategy of the modern genetic technology and the traditional breeding method is combined.

The invention provides a construction method of a recombinant canine distemper virus strain for expressing mink parvovirus VP2 protein, and relates to the technical field of genetic engineering. According to the construction method, a mink canine distemper virus attenuated vaccine CDV3 strain is adopted as a basis, a mink parvovirus VP2 gene is cloned into canine distemper virus genome cDNA, and a recombinant canine distemper virus rCDV3-mVP2 strain capable of simultaneously expressing CDV protein and MEV VP2 protein is obtained through virus reverse genetic technology rescue. A candidate vaccine strain is provided for research and development of a mink canine distemper and parvovirus enteritis bigeminal recombinant live vaccine.

The present invention belongs to the field of plant genetic technology, and discloses a new use of S-adenosyl-L-homocysteine hydrolase gene. The present invention reveals that the S-adenosyl-L-homocysteine hydrolase gene Gomus.A12G023400 can be used to improve the fiber quality of cotton or for molecular marker-assisted breeding of high-strength fiber cotton varieties. The technical solution of the present invention provides a new way for molecular marker-assisted improvement breeding of cotton fiber quality.

The present invention relates to the field of biological vaccine research and development technology, and in particular, to a VSV vector-based highly pathogenic H5 subtype avian influenza inactivated vaccine and a method for preparing same. The method comprises: using a vesicular stomatitis virus with the deletion of the surface envelope protein gene as a vector, inserting the NA gene and the HA gene of an H5 subtype highly pathogenic avian influenza virus strain between the M gene and the L gene of the vector in the form of an expression cassette, and acquiring a recombinant virus by means of rescue via reverse genetic technology; inoculating the recombinant virus into susceptible cells, culturing for replication and proliferation, and then harvesting a viral solution; inactivating the viral solution to acquire an inactivated viral solution; and adding the inactivated viral solution to a pharmaceutically or veterinarily acceptable carrier, excipient, medium, or adjuvant to acquire an inactivated vaccine. The provided H5 subtype avian influenza inactivated vaccine can induce a high hemagglutination inhibition titer of avian influenza virus on day 21 after chick immunization. The provided inactivated vaccine has good safety and immunogenicity.

The invention provides a specific InDel molecular marker of a key gene OsSPO11-1 for rice apomixis control and application of the specific InDel molecular marker, and belongs to the technical field of crop molecular heredity. The invention provides a 9bp deletion mutant of a second exon of a rice fertility control gene OsSPO11-1 at the 162-170bp position in a CDS (Coding Sequence) sequence. The mutation causes the deletion of three key amino acids His-Arg-Tyr in the amino acid coding sequences from the 54th site to the 56th site. In fertility expression, the homozygous type of the mutant shows the characteristic of complete sterility, and the heterozygous type of the mutant keeps complete fertility. In order to accurately identify the mutation, the invention specifically develops a specific molecular marker, as shown in SEQ ID NO.7. The invention also provides a method for identifying the mutation. Through single PCR amplification, the marker can accurately distinguish a wild type, a heterozygous type and a mutant type, and the detection result is completely consistent with the results of sequencing analysis and phenotype observation.

The invention belongs to the technical field of molecular biology, and particularly relates to a preparation method and application of virions. According to the invention, a reverse genetic technology is utilized to construct a rift valley heat replication defective virus-like particle for visual tracking of virus infection dynamics. The virion is only subjected to amplification passage in NSm-Gn-Gc expression cells, only one round of infection occurs in other cells, amplification passage cannot be performed, the infectivity of the rift valley fever virus is reduced, and the operation difficulty and the exposure risk are reduced. According to the quantum dot-labeled particles, real-time visualization of infection dynamics is realized, and dynamic characteristics of virus invasion links are accurately quantified. The content of the invention can be expanded to animal models for tracking tissue organs that a single virion initially targets when invading a host in a non-replication state. A novel technical platform is established for exploring an infection replication mechanism of the rift valley fever virus, and an important reference is provided for tracking research of other high-pathogenicity viruses.

The invention discloses an influenza H5 subtype recombinant virus based on a VSV carrier, an inactivated vaccine and a preparation method of the influenza H5 subtype recombinant virus, and relates to the technical field of research and development of biological vaccines. The genome of the recombinant virus comprises a VSV genome without a surface envelope protein gene, and an NA gene and an HA gene of an H5 subtype avian influenza virus strain; the NA gene and the HA gene are located between an M gene and an L gene of a VSV genome, and the NA gene and the HA gene are connected in series in the 5 '-3' direction. The recombinant virus rescued by the reverse genetic technology can effectively express the NA antigen and the HA antigen of the H5 subtype avian influenza virus. The inactivated vaccine provided by the invention comprises the inactivated recombinant virus and a vaccine adjuvant, the inactivated recombinant virus loses replication ability, the NA protein and HA protein of the inactivated virus can still play an immune role under the assistance of the specific adjuvant, high-level avian influenza virus hemagglutination inhibition titer can be induced to be generated after a mouse is immunized, and the inactivated vaccine can be used for treating avian influenza virus hemagglutination. The method has the advantage of high safety.

The present invention belongs to the field of plant genetic technology, and discloses a method for discovering genes related to cotton fiber strength and its application. The present invention reveals a method for discovering genes related to cotton fiber strength based on lncRNA analysis by using near-isogenic lines with different fiber qualities, and specifically applies this method to obtain three cotton fiber strength genes, namely Gomus.D05G015100, Gomus.A05G281300, and Gomus.A10G226800; these three genes can be used to improve the fiber quality of cotton or for molecular marker-assisted breeding of high-strength fiber cotton varieties. The technical solution of the present invention provides a new approach for molecular marker-assisted improvement breeding of cotton fiber quality.

The present invention relates to the fields of bioinformatics and genetics, and more particularly to a method and apparatus for detecting introgression between biological populations. The method comprises obtaining genetic information from at least three biological populations, scanning the genetic information, and determining the location and number of target genomic regions exhibiting introgression within the genetic information; and determining whether introgression exists between the biological populations based on the location and number of the target genomic regions within the genetic information. Compared to existing methods, the method of the present invention offers higher detection efficiency.

The invention belongs to the technical field of reproduction and heredity, and relates to application of SgI-52, in particular to application of SgI-52 in preparation of a reagent for screening high-quality sperms and a screening method. The invention finds that the SgI-52 and the PRDX6 are selectively combined with poor-quality sperms, based on the finding, a flow cytometry is applied to sort and screen out sperms which are combined with the SgI-52 and highly express the PRDX6 by plasma membranes according to whether the SgI-52 is combined with the PRDX6 on the sperms in a semen sample or not, so that the sperms which are not combined with the SgI-52 and have higher quality are optimized, and finally, the semen sample with high-quality sperms is obtained. The method has the advantages that the success rate of the assisted reproduction technology is increased, and the important function and mechanism in asthenospermia sperm screening provide theoretical basis and key technology for sperm optimization, so that the technical support is provided for the male infertility technology.

The invention belongs to the technical field of virus reverse heredity, and particularly relates to a construction method and application of a Newcastle disease virus efficient recombinant reverse heredity system based on a CPER technology. The construction method comprises the following steps: carrying out CPER assembly on the Newcastle disease by using a linker fragment for coding Pol-I, preparing an NP / P / L helper plasmid, verifying a recombinant virus after co-transfecting cells, and the like. By using the reverse genetic system prepared by the invention, the need for molecular cloning of bacteria can be omitted, so that various recombinant viruses with mutated and / or inserted exogenous genes can be designed and generated in a short time. The recombinant Newcastle disease virus prepared by the method lays a good foundation for development of related novel Newcastle disease vaccines. Meanwhile, the reverse genetic system can generate more viruses and overcome the problem of low transfection efficiency, so that the reverse genetic system has a wide application prospect.

The invention relates to construction and application of a recombinant RSV infectious clone, Long-BAC is taken as a basic skeleton, an F gene of a Long strain is respectively replaced with an F gene of an RSV A subtype global preponderant epidemic genotype ON1 and an F gene of a B subtype BA9 genotype, and two attenuated recombinant virus strains are obtained through rescue by a reverse genetic technology. The technical method disclosed by the invention has the advantages of rapidness and accuracy, the recombinant virus containing the epidemic genotype F gene can be successfully constructed, and the recombinant virus provides a scientific and technical platform for researching the influence of F protein variation on virus transmission capacity, host adaptability and pathogenicity and for neutralizing and escaping evaluation of vaccines and antibody drugs.

The invention discloses a highly pathogenic H5 subtype avian influenza inactivated vaccine based on a VSV carrier and a preparation method thereof, and relates to the technical field of research and development of biological vaccines. The preparation method of the inactivated vaccine comprises the following steps: by taking a vesicular stomatitis virus without a surface envelope protein gene as a carrier, inserting an NA gene and an HA gene of an H5 subtype highly pathogenic avian influenza virus strain between an M gene and an L gene of the carrier in the form of an expression box, and rescuing through a reverse genetic technology, thereby obtaining the inactivated vaccine. Inoculating the recombinant virus to a susceptible cell for culturing, and replication and proliferation to obtain a virus solution; carrying out inactivation treatment on the virus solution to obtain an inactivated virus solution; and adding a pharmaceutically or veterinary acceptable carrier, an excipient, a medium or an adjuvant into the inactivated virus liquid to obtain the inactivated vaccine. According to the H5 subtype avian influenza inactivated vaccine provided by the invention, chicken immunized with the H5 subtype avian influenza inactivated vaccine can be induced to generate high-level avian influenza virus hemagglutination inhibition titer after 21 days. The inactivated vaccine provided by the invention has high safety and good immunogenicity.

The invention discloses a genotype filling optimization method for spruce norvegensis and a reference panel, and belongs to the technical field of forest heredity. The method comprises the following steps: constructing a haplotype reference panel based on large-scale picea norvegensis re-sequencing data; acquiring low-density genotype data; carrying out genotype filling by using IMPUTE5 software and integrating a genetic map of the picea norvegensis; and carrying out quality control on a filling result, and reserving a high-quality SNP site with Rsqsoft greater than or equal to 0.8. According to the method, key parameters and optimal configuration influencing filling accuracy are evaluated and determined for the spruce norveyi system for the first time, and the technical bottleneck that the species are lack of a reliable filling process is solved. Through the method and the special reference panel provided by the invention, high-quality whole genome filling data can be stably and efficiently obtained, and key technical support is provided for genetic research and breeding practice of picea norvegensis.

The invention discloses a method for rapidly constructing a broad-spectrum cross protection type IBV nested epitope vaccine. According to the method, computer simulation docking and in-vitro immunological screening are combined, conservative and effective antigen epitopes are rapidly and accurately identified from a large number of strain sequences, the antigen epitopes are embedded into a virus genome S1 structural domain on the basis of a reverse genetic technology, and therefore the nested epitope vaccine strain rSczy3C100-S1T is obtained. The problems that an existing IBV vaccine is narrow in immune protection spectrum, poor in cross protection and long in research and development period are solved, candidate vaccines aiming at emerging strains can be constructed in a short time, and the vaccine development efficiency and broad-spectrum protection capacity are remarkably improved.