41 results about "Fugu rubripes" patented technology

The invention discloses a preparation method of a recombinant fugu rubripe interferon gamma protein. The method has advantages of simple operation, low cost, high purity of the recombinant protein and high yield, and is suitable for industrial production. The preparation method comprises steps of: extracting the total RNA of fresh liver of fugu rubripes and performing inverse transcription to synthesize cDNA of a first strand; performing RT-PCR amplification by adopting the cDNA of the first strand as a template, with the upstream primer being N[co] I:5'-CATGCCATGGGCTCCTACATCCCAGTAGAAATG-3', and with the downstream primer being X[ho] I:5'-CCGCTCGAGATGACCTCGAATCGACATCT-3'; establishing a clone vector, expressing the vector, transferring into an expression bacterial, and cultivating the bacterial strain to obtain strain; subjecting the collected strain to enlarging cultivation, inducing with IPTG and collecting the bacterial strain; and smashing the bacterial strain, lysing, purifying, and collecting purified liquid.

The invention discloses a temperature and light regulation and control method for out-of-season breeding of fugu rubripes parent fishes and application, and belongs to the technical field of factory-like circulating water cultivation of the fugu rubripes parent fishes. The breeding period of the parent fish of fugu rubripes is concentrated from March to May, and the production of fries is limited by seasons. By simulating the illumination period and the annual change period of the water temperature, the change rate of the annual period is accelerated from one time of 12 months to one time of 6 months, gonad development of the parent fish is promoted to be advanced, and breeding and egg laying in winter are achieved. The fry obtained by out-of-season breeding is easy to be subjected to low-temperature masculinization induction.

The invention relates to an in-vitro construction method and application of a fugu rubripes ovarian cell line. The construction method comprises the steps of (1) preparing a primary cell culture solution; (2) performing primary culture; (3) performing subculture; and (4) separating cell lines with different forms. The application comprises application of the fugu rubripes ovarian cell line to multiplication and expanding propagation of two typical fish viruses and application of the fugu rubripes ovarian cell line to transgenic research. According to the in-vitro construction method, an ovary is used as a culture and line construction object to build a biological platform for research on extraction and utilization of tetrodotoxin and safe detoxification treatment of cultured globefish, so that the conventional aquaculture industry and the emerging bio-pharmaceutical industry can be well served.

The invention discloses a SNP (Single Nucleotide Polymorphism) primer for screening fugu rubripes seedlings and a screening method, which are capable of saving manpower and material resources, lowering seed screening cost and guaranteeing excellent characters of the filial generation. The sequence of the SNP primer is as shown in SEQ ID NO.1 and SEQ ID NO.2. The seedling screening method comprises the following steps: extracting the genomic DNA of to-be-detected fugu rubripes; taking the acquired genomic DNA as a template; performing PCR amplification with upstream primer and downstream primer, thereby obtaining a PCR product; sequencing and genetically typing the obtained PCR product; screening the fugu rubripes with DNA sequence as shown in SEQ ID NO.3 and the No.591 locus gene started from 5' end being homozygosis TT.

The invention discloses a method for preparing a tetrodotoxin positive quality control sample. The method comprises the steps of selecting muscular tissues from fresh fugu rubripes, homogenizing, and measuring the content of tetrodotoxin in the muscular tissues to verify whether the muscular tissues contain the tetrodotoxin; if the muscular tissue is a positive sample, diluting the positive sample containing toxin with a negative sample and a dispersant, mixing uniformly, continuously measuring the content of the toxin in the sample, packaging the sample after the sample meets requirements, preserving at an appropriate temperature, and performing uniformity verification and stability analysis on the sample; if the muscular tissue is a negative sample, adding a tetrodotoxin standard solution into the negative samples, adding a dispersant, mixing uniformly, measuring the content of the tetrodotoxin in the sample, packaging the sample after the sample meets requirements, preserving at an appropriate temperature, and performing uniformity verification and stability analysis on the sample. The quality control sample is simple to operate, and has enough stability and uniformity and long preservation time, so that the quality control sample can be used for laboratory quality control.

The invention discloses a method for preparing a marine filariasis vaccine of redfin puffer. A single marine filariasis is picked and inoculated in L-15 complete medium for purification and cultivation, and then enlarged and cultivated, and 100 μl / L of Foreria is added. Marin killed the larvae, centrifuged at 4000rpm in a low-speed centrifuge for 10 minutes to collect the precipitate, added L‑15 medium to prepare an inactivated larvae liquid with a density of 2 million / ml, collected in a 1.5ml centrifuge tube, and stored at ‑80°C. Before immunization, thaw the dead larvae on ice, crush them with electromagnetic waves, mix them with the same amount of Freund's adjuvant or Freund's incomplete adjuvant, and shake the mixture until it is in the form of emulsion, which is to prepare the marine tail silk of puffer puffer Insect vaccines. Intraperitoneal injection can improve the immunity of pufferfish against marine filariasis.

The invention discloses a method for inducing masculinization of fugu rubripes at a low temperature. The method comprises the following steps of: performing artificial induced spawning and insemination on healthy male and female fishes which are high in fatness and undergo good gonad development; hatching fertilized eggs under the water temperature condition of 20 DEG C for 45-75 days; and putting into water of 15-17 DEG C, and culturing for 35-65 days. Due to the adoption of the method, male fugu rubripes fries can be produced rapidly, the male rate in the fries is 65-80 percent, the average male rate is 75 percent, ultraviolet ray irradiation is not required, sex hormone is not fed, and complex operation processes are avoided; and the method is low in cost, is easy to operate, and has contribution to mass production.