12 results about "Paralichthys" patented technology

The invention belongs to the field of biotechnology and aquaculture, and particularly relates to a method for improving hypoxia resistance of pelteobagrus vachelli based on hypoxia induction of liver extracellular vesicles of the pelteobagrus vachelli. The method comprises the following steps: extracting high-purity extracellular vesicles EVs in liver tissues of pelteobagrus vachelli subjected to low-oxygen treatment by adopting a method of combining differential centrifugation and high-precision iodixanol density gradient centrifugation; diluting the extracted EVs to 2mu g / mu L by using PBS (Phosphate Buffer Solution) to prepare an EVs suspension; an intraperitoneal injection mode is adopted, the EVs suspension is injected into the body of the pelteobagrus vachelli, and after injection is conducted for 24 hours, the hypoxia resistance of the pelteobagrus vachelli is evaluated through a suffocation point detection test. The invention opens up a brand new path for improving the hypoxia resistance of the pelteobagrus vachelli, provides a new idea and a new method for efficiently enhancing the hypoxia resistance of economic fishes in the field of aquaculture, is expected to play an important role in actual aquaculture production, and assists the sustainable development of the aquaculture industry.

The application discloses a tandem antigen epitope peptide of G protein of Paralichthys olivaceus rhabdovirus and application thereof, and belongs to the field of fish molecular immunology. The amino acid sequence of the tandem antigen epitope peptide is shown as SEQ ID NO:1. The preparation method of the application comprises the following steps: (1) first, analyzing the structural characteristics of HIRRV-G protein and predicting B cell antigen epitopes, and synthesizing the antigen epitopes with advantages based on the prediction results; (2) then, screening candidate peptide segments with high affinity through enzyme-linked immunosorbent assay; (3) sequentially connecting the high-affinity peptide segment sequences meeting the requirements by using a GPGPG linker, and cloning the connected sequences into a pET-28a prokaryotic expression vector, so that the tandem antigen epitope peptide is obtained after induced expression. Compared with the full-length G protein, the tandem antigen epitope peptide has smaller molecular weight, stronger stability and stronger hydrophilicity; the tandem antigen epitope peptide can induce a large amount of specific antibodies in fish bodies, and significantly reduces the mortality of fish infected by the virus. The tandem antigen epitope peptide can be used for the development of HIRRV diagnostic reagents and subunit vaccines.

The invention belongs to the technical field of biology, and particularly relates to a pharmaceutical composition for inducing gonad exhaustion of paralichthys olivaceus and application of the pharmaceutical composition. The pharmaceutical composition comprises docetaxel and cis-platinum, the mass ratio of docetaxel to cis-platinum is 1: 1, and the total concentration of docetaxel and cis-platinum is 100-200 mu g / mL. By optimizing the medicine proportion and the administration scheme, the problems of low survival rate and large injury of fish bodies in the existing gonad exhaustion method are solved, and efficient and stable preparation of the paralichthys olivaceus germline stem cell transplantation receptor can be realized. By controlling the ratio of docetaxel to cis-platinum, the administration concentration, the injection frequency, the action period and the like, the invention aims to solve the problems of insufficient exhaustion effect, low fish survival rate or serious gonad injury and the like in the existing receptor preparation, and provides reliable technical support for establishing a stable and efficient paralichthys olivaceus germline stem cell transplantation receptor system.

The application provides a double-strand DNA vaccine for late Edwardsiella tarda disease, which can significantly induce Th1 type immune bias, enhance fish cell immune response, improve the resistance to intracellular bacterial infection, and has the advantages of safety and high efficiency, easy preparation, strong adaptability for on-site promotion and the like. The double-strand DNA vaccine uses a fusion fragment of an immune adjuvant IL-12, which is constructed by connecting a nucleic acid fragment coding a subunit 12p35 with an amino acid sequence of SEQ ID NO:1 and a nucleic acid fragment coding a subunit 12p40 with an amino acid sequence of SEQ ID NO:3 through a flexible connecting peptide linker. The double-strand DNA vaccine realizes the synergistic expression of OmpC antigen and IL-12 adjuvant, enhances immunogenicity, and solves the problem of insufficient immune effect of a traditional DNA vaccine; and the vaccine has no obvious toxic side effect on fish bodies, is not dependent on the use of antibiotics, is green and environmentally friendly, and has a good industrialization prospect.

The invention provides application of an overexpressed ifitm3 gene in promoting in-vitro steady state maintenance of oogonial stem cells of paralichthys olivaceus, and belongs to the technical field of gene engineering. The overexpression vector containing the ifitm3 gene is constructed and transfected to the oogonial stem cells of the paralichthys olivaceus, so that the in-vitro proliferation activity and migration ability of the stem cells can be remarkably enhanced, and the undifferentiated state and the stemness characteristic of the stem cells are effectively maintained. The technology provides a key means for in-vitro long-term culture, directional induced differentiation and excellent germplasm resource preservation of paralichthys olivaceus oogonial stem cells, and has important application value in fish reproductive biology research, gene editing breeding and culture industries.

The invention belongs to the technical field of genetic breeding, and particularly relates to a preparation method of a novel high-temperature-resistant and fast-growing paralichthys olivaceus triploid strain. According to the method, high-temperature resistance and body weight are used as screening indexes, a high-temperature-resistant commercial line and a fast-growing commercial line of paralichthys olivaceus are constructed by using the technologies of population breeding, gynogenesis and the like, then the commercial lines are used as male and female parents, mature sperms and eggs are collected for insemination, triploid induction is performed, and high-temperature resistance and growth traits are measured, so that the high-temperature-resistant commercial line of paralichthys olivaceus is obtained. And obtaining a triploid new strain with two characters of high temperature resistance and fast growth at the same time. According to the method, the triploid paralichthys olivaceus fries with high-temperature-resistant and fast-growing characters can be efficiently induced and generated in batches, and high practicability is achieved.

This invention discloses a method for the zonal symbiotic aquaculture of striped sawfish and turbot in a recirculating aquaculture system, belonging to the field of aquaculture technology. The method involves dividing the aquaculture pond into upper and lower zones within a unified recirculating aquaculture system using permeable mesh screens. Striped sawfish are introduced into the upper and middle layers, while turbot are introduced into the lower zones. The water temperature is controlled at 20-24℃, with a photoperiod of 12L:12D, with low light intensity in the upper zone and shading in the lower zone. The upper zone is fed floating extruded feed, while the lower zone is fed sinking soft pellet feed, with staggered feeding times. Water quality is controlled with dissolved oxygen ≥6mg / L, ammonia nitrogen <0.1mg / L, nitrite <0.1mg / L, and pH 7.8-8.2. Regular monitoring and zonal harvesting are conducted. This invention utilizes physical zoning and complementary ecological niches to avoid interspecies competition and interference, enabling two high-value fish species to coexist efficiently in the same water body. This significantly increases yield and economic benefits per unit water volume, reduces aquaculture risks, saves energy and reduces consumption, and allows for precise management, making it suitable for the promotion and application of factory-scale recirculating aquaculture.

The application provides an antigen epitope peptide and a paralichthys olivaceus mucin Muc13 specific antibody prepared by the antigen epitope peptide, determines a B cell epitope from the paralichthys olivaceus Muc13 protein through antigen epitope screening, and the sequence is PGTTEEPDPTAGSPP. After the epitope is coupled with a KLH carrier protein, the epitope is used for immunizing New Zealand white rabbits, and a rabbit-derived anti-paralichthys olivaceus Muc13 specific antibody is successfully obtained. The antibody can specifically recognize the Muc13 protein in paralichthys olivaceus mucosa tissue and Muc13 positive mucous cells, and provides a key detection means for in-depth exploration of the function mechanism of mucous cells in the fish mucosa immune response process.

The application provides an antigen epitope polypeptide and a paralichthys ovatus mucin Muc1 specific antibody prepared by the antigen epitope, a B cell epitope from the paralichthys ovatus Muc1 protein is identified through antigen epitope screening, and the amino acid sequence is SEQ ID NO:1. After the epitope is coupled with a KLH carrier protein, a New Zealand white rabbit is immunized, and a rabbit anti-paralichthys ovatus Muc1 polyclonal antibody is successfully prepared. The antibody can specifically recognize the Muc1 protein in paralichthys ovatus mucosa tissue and positive mucous cells, and provides an important detection tool for studying the function mechanism of mucous cells in fish mucosal immunity.

According to the bicistronic DNA vaccine for edwardsiella tarda disease of paralichthys olivaceus, provided by the invention, the bicistronic DNA vaccine can remarkably induce Th1 type immune bias, enhance immune response of fish cells and improve the resistance to intracellular bacterial infection, and has the advantages of being safe, efficient, easy to prepare, high in field popularization adaptability and the like. A fusion fragment of an immunologic adjuvant IL-12 is used in the bicistronic DNA vaccine, and the bicistronic DNA vaccine is constructed by connecting a nucleic acid fragment of a subunit 12p35 of which the coding amino acid sequence is SEQ ID NO: 1 and a nucleic acid fragment of a subunit 12p40 of which the coding amino acid sequence is SEQ ID NO: 3 through a flexible connecting peptide joint. According to the bicistronic DNA vaccine disclosed by the invention, the synergistic expression of the OmpC antigen and the IL-12 adjuvant is realized, the immunogenicity is enhanced, and the problem of insufficient immune effect of the traditional DNA vaccine is solved; and the vaccine has no obvious toxic or side effect on the fish body, does not depend on antibiotics, is green and environment-friendly, and has a good industrialization prospect.

The application provides a method for breeding a Paralichthys olivaceus clone improved strain, and the method comprises the following steps: preparing a high-purity Paralichthys olivaceus gynogenesis diploid, preparing a Paralichthys olivaceus double haploid (DH), preparing a double clone hybrid strain, preparing a recombination double haploid (rDH), cultivating the Paralichthys olivaceus recombination double haploid (rDH), and preparing and cultivating the clone improved strain. The Paralichthys olivaceus clone improved strain with fast growth and high reproductive capacity is bred, and the problem of practical application of fish double haploid breeding can be solved.

The invention belongs to the technical field of fish genetic breeding and genetic engineering, and particularly relates to a rapid preparation method of a gene editing fish homozygous mutant based on a gynogenesis technology. According to the method, F0 or F1 generation female marine fishes such as paralichthys olivaceus obtained through gene editing operation serve as female parents, mature ova of the female marine fishes are collected, and the ova are activated through sperms subjected to genetic material inactivation treatment; then, chromosome doubling is induced in a cleavage inhibition mode, and the mitotic gynogenesis diploid is obtained. And further screening the filial generation obtained by the steps to obtain the mutant with homozygous genotype. The method is simple and convenient to operate, the period of obtaining the homozygous mutant can be remarkably shortened, the breeding efficiency is improved, and the method has a good application prospect.