77 results about "Genetically modified fish" patented technology

The present invention relates to the method and use of fluorescent proteins in making purple transgenic fluorescent fish. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing. Thus, new varieties of ornamental fish of different fluorescence colors from a novel source are developed.

The present disclosure provides methods of producing enucleated cells by photoablation. Such enucleated cells may be used as recipient cells for Somatic Cell Nuclear Transfer and cloning. The nuclear donor and / or enucleated recipient cells may be any fish cells, such as zebrafish, koi, or medaka fish cells. Such methods may be used to efficiently produce transgenic fish including by way of example zebrafish, koi, and medaka fish.

In accordance with the present invention, there are provided humanized fish insulin genes. Humanized insulin the present invention encode human insulin alpha and / or beta chains while using fish-preferred codons and regulatory sequences. These humanized genes are thus expressible in fish islet cells. Also provided are transgenic fish having islet cells containing and capable of expressing humanized insulin genes. These islet cells (Brockmann Bodies) can be xenotransplanted into subjects having diabetes. In this manner normoglycemia can be achieved in the recipient of the islets.

The present invention relates to transgenic green ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing.

The invention discloses a vector for efficiently labeling zebra fish PGC, and a preparation method and a use of transgenic fish. A Gal4 / UAS transcription activation system is utilized and mRFP is used as a report gene to realize an inductive gene expression regulation technology in the primordial germ cells of zebra fish. The preparation method of the transgenic fish comprises the following steps: 1, respectively constructing activation transgenic line Tg (kop:KalTA4) and an effect transgenic line Tg (UAS:mRFP); and 2, hybridizing Tg (kop:KalTA4) male fish with Tg (UAS:mRFP) female fish to obtain the transgenic fish for efficiently labeling the primordial germ cells of zebra fish. The transgenic fish for efficiently labeling the primordial germ cells of zebra fish can be widely applied to the fish bioengineering.

The present invention relates to the method and use of fluorescent proteins in making purple transgenic fluorescent fish. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing. Thus, new varieties of ornamental fish of different fluorescence colors from a novel source are developed.

The present invention relates to transgenic green ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing.

The present invention relates to transgenic fluorescent orange ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing.

The present application disclosed a method for preparing a sterile transgenic fish, comprising constructing antisense RNA expression vector of salmon-type gonadotropin-releasing hormone, introducing the recombinant DNA fragment into carp oosperm by microinjection, and screening the sterile transgenic fish by Polymerase Chain Reaction and radioimmunoassay, wherein the expression vector comprising a promoter of carp beta actin (β-actin) gene, a complementary DNA fragment of antisense salmon-type gonadotropin-releasing hormone (sGnRH) gene from carp with 323 bp as a target gene comprising sGnRH decapeptide, the coding region of gonadotropin-releasing hormone associated peptide and 3′ non-coding sequence, and 3′ flanking sequence of grass carp growth hormone gene as a stop sequence. The method of the present invention is easy and convenient for operation which provides a basis for providing a technical platform of general applicability for solving the hereditary and ecological safety problems of transgenic fishes.

A method of inducing male and / or female infertility in a genetically modified (GM) fish is disclosed. Also disclosed is a method of generating an infertile GM fish with a phenotype and / or genotype of interest. The method involves generation of a GM fish whose genome comprises a foreign transgene operably linked to a fish gonad-specific promoter selected from the group consisting of an ovary-specific promoter and a testis-specific promoter. The transgene comprises a suicide gene selected from the group consisting of a reductase and a photosensitizer, in which the reductase is operably linked to a reporter gene. Infertility of the GM fish is induced if the GM fish expressing the reductase is treated with an effective amount of a reductase-activated cytotoxic prodrug or if the GM fish expressing the photosensitizer is treated with light irradiation.

The invention aims at providing a transgenic method of a marine flounder. The transgenic method of the marine flounder comprises the following steps of: transferring plasmids with exogenous genes into pubescent testis of a milter; and collecting seminal fluid of the transgenic milter and fertilizing with mature eggs of a hen, so as to transfer the exogenous gene into the mature eggs through sperms. The method avoids the problem that lots of embryos are dead due to micro-injection to the oosperm. By adopting the method, the exogenous gene can be effectively transferred into the oosperm of the flounder; the integration of the exogenous gene integrated in a transgenic fry is 12 to 21%; a novel method is provided for the study on gene transfer of the flounder; the problem that the transgenosis is hardly carried out on the oosperm of the flounder by micro-injection is solved; and novel technical means are provided for breeding in the flounder genetic engineering.

The invention discloses a method for breeding Macropodus opercularis with a transferred green fluorescent protein gene by using piggyBac transposons, which comprises the following steps of: firstly, constructing piggyBac transposons with green fluorescent protein genes and a helper plasmid capable of providing transposase; and secondly, transferring the two transposons into a germ cell of the Macropodus opercularis by using microinjection technology, and according to the transposons characteristics of the piggyBac transposons, introducing the green fluorescent protein genes into a genome of the Macropodus opercularis so as to make the gene inherited and expressed stably. An individual of transgenic Macropodus opercularis can be screened by means of the fluorescence-labeled gene, and the Macropodus opercularis with the transferred green fluorescent protein gene is more worthy of appreciation and represents a new variety of Macropodus opercularis with specific function. In addition, it is the first time for the piggyBac transposons to be used in researches on transgenic fish, and thus, the research space for transgenic aquatic animals is widened.

The invention discloses a method for constructing a zebrafish kidney progenitor cell marker transgenic line. The method comprises the following steps: a gene knock-in method is used to construct a transgenic fish line of Six2a:GFF in a zebrafish, wherein the fish line hybridizes with UAS:GFP, UAS:Dendra2-NTR to specifically label kidney progenitor cells in the kidney or delete kidney progenitor cells, as well as overexpression and inhibition of the genes are carried out in kidney progenitor cells. The line can be used for high-throughput drug screening to provide the method to create kidney progenitors in human kidneys.

The invention discloses a larimichthys crocea interferon regulatory factor IRF7 starter, a nucleic acid builder, a cell and a preparing method and application of the larimichthys crocea interferon regulatory factor IRF7 starter, the nucleic acid builder and the cell. The nucleotide sequence of the larimichthys crocea interferon regulatory factor IRF7 starter is shown as Seq ID No.1 or a nucleotide sequence complementary to Seq ID No.1. The invention further protects application of the starter or the nucleic acid builder or a carrier or a reconstitution cell or fish or a mammal containing the starter or the nucleic acid builder or the carrier or the reconstitution cell in regulating target gene expression or transgenic fish.

The invention provides a solution for prolonging the opening time of fertilization holes of fish eggs and a method for introducing foreign genes by using the fertilization holes, and belongs to the technical field of research and production of transgenic fish. The solution uses water as a solvent, and each liter of the solution comprises 6 g of sodium chloride, 0.15g of potassium chloride, 0.1g ofcalcium chloride, 0.1g of sodium bicarbonate, 0.1g of sodium dihydrogen phosphate and 1g of glucose. The solution can prolong the opening time of the fertilization pores of the fish eggs, so that theforeign genes can conveniently directly enter the fish eggs.

The present invention relates to fish with increased unsaturated fatty acid content by transfer of a fatty acid desaturase gene thereinto, and a production method of these fish with increased unsaturated fatty acid content. The fatty acid desaturase gene is one or more members selected from among Δ4 fatty acid desaturase gene, Δ5 fatty acid desaturase gene and Δ6 fatty acid desaturase gene, and it is preferable that these genes originate in a freshwater fish. It is preferable that the expression of a fatty acid desaturase gene originating in a freshwater fish is promoted by a β-actin promoter originating in medaka. By modifying the fatty acid metabolic pathway of fish, problems to be solved by the present invention may be to provide fish having a high vitality and a tolerance to handling and low temperature, which will enable the use of a vegetable fat or animal fat as a formula feed. Such vegetable fat is less expensive and can be easily controlled in qualities, thereby largely contributing to the reduction of cost and labor in fish nursery sites.

The invention discloses a lentivirus method for preparing IGF2b transgenic fish. The method is characterized in that: the production of the transgenic fish is carried out by establishing lentivirus vectors containing target genes; the vectors contain GFP reporter genes which can be used for testing the transgene effect. The method provided by the invention is used for producing lentivirus transgenic fish. The survival rate of the transgenic fish is 12.2%.

The invention provides methods for producing biofuel from algae, that use fish which have a high capacity of producing and / or accumulating lipids to harvest algae from an algal culture. The invention also provides methods for growing fish that result in a high lipid content. The invention also provides methods for creating fish that have a high capacity of producing and accumulating lipids by breeding and / or recombinant DNA techniques. Also included are transgenic fish that have a higher lipid content than wild-type fish.

The invention relates to a device for automatically micro-injecting fish embryonated eggs and automatically sorting juvenile fishes. The device comprises a sampling part and an optionally-injecting part and / or a sorting part, so that the transgenic fish can be effectively built, automatic injection of the fish embryonated eggs is realized, and the automatic sorting of the juvenile fishes is realized.

The present invention relates to expression control sequences of a vertebrate liver fatty acid binding protein (L-FABP) gene that, when operably linked to a reporter (e.g., a heterologous reporter, such as the green fluorescent protein (GFP)), directly express the reporter in a fashion that mimics the liver-specific development of the L-FABP gene in the vertebrate. Also disclosed is transgenic fish, such as a transgenic zebrafish, whose cells comprises at least one genomically integrated copy of a recombinant construct comprising such an expression control sequence, operably linked to a reporter sequence, so that the expression of the reporter is liver-cell specific, both spatially and temporally during development.

It is intended to utilize a vegetable fat, which is less expensive and can be easily controlled in qualities, thereby largely contributing to the reduction of cost and labor in fish nursery sites and, moreover, enabling the production of fish fry having a high vitality and being tolerant to handling and low temperature without resort to fish oil containing EPA and DHA as a formula feed for fish cultivation. Namely, fish with an increased unsaturated fatty acid content in which a fatty acid desaturase gene is expressed due to the transfer of the fatty acid desaturase gene thereinto. The fatty acid desaturase gene is one or more members selected from among Delta4 fatty acid desaturase gene, Delta5 fatty acid desaturase gene and Delta6 fatty acid desaturase gene, and it is preferable that these genes originate in a freshwater fish. It is preferable that the expression of a fatty acid desaturase gene originating in a freshwater fish is promoted by a beta-actin promoter originating in killifish.

The invention discloses a continuously activated growth hormone receptor gene of fishes, and a preparation method and application thereof. The preparation method comprises the following steps of: respectively performing polymerase chain reaction (PCR) amplification on growth hormone receptor (GHR) signal peptide, a c-Jun leucine zipper region, a GHR transmembrane region and an intracellular region of carp, and combining another turn of bridge-PCR amplification to obtain a continuously activated GHR recombinant gene segment; and cloning the obtained recombinant gene into an expression vector pCA-AT to obtain a recombinant gene vector pCA-SJG-AT for gene transfer. A protein encoded by the gene has the amino acid sequence shown as SEQIDNO:1; and a separated recombinant gene has the nucleotide sequence shown as SEQIDNO:2. The application of the recombinant gene in quick growth of transgenic fish has the advantages that: (1) the activity of the recombinant GHR protein for promoting a signal channel is high, and the efficiency of the continuously activated GHR effect is more than 3 times higher than that of GH effect; and (2) the transgene of the continuously activated GHR gene has obvious growth promotion effect, and the transgenic fish has a growth speed which is over 50 percent higher than that of contrast fish.

The present invention relates to transgenic orange ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing.

The present invention relates to a transgenic fish having at least one genomically integrated expression cassette containing a 5′-regulatory nucleotide sequence responsive to hormones, particularly estrogenic hormones, connected in a functional manner upstream of a nucleotide sequence encoding a reporter protein. The present invention further relates to methods of using the transgenic fish for various purposes, including, for example: (1) identifying estrogenic endocrine disruptors; (2) monitoring estrogen-like activity of test samples; (3) identifying anti-estrogenic endocrine disruptors; and (4) investigating the effects of endocrine disruptors on liver regeneration. Expression cassettes, host cells, and transgenic cells of aquatic animals are also disclosed.

The invention discloses a quick identification method and application of a transgenic fish homozygote. The method comprises the following steps: A. cutting about 0.5-1cm<2> of transgenic fish tail fin tissues, and extracting DNA (deoxyribonucleic acid); B. detecting that the transplanted gene is positive by conventional PCR technology; and C. analyzing the genotype of the transgenic fish of which the transplanted gene is positive by an SYBRGreenreal-timePCR 2<DELTADELTACt> method, comparing the integrated relative number of copies of the transplanted gene in the homozygote transgenic fish and hemizygote transgenic fish, identifying and screening out the transgenic homozygote, and further verifying by a test-mating experiment. The method is easy to implement and simple to operate, and is quick and accurate. The identification and screening of the transgenic fish homozygote have important meanings for establishing the transgenic fish pure line and accelerating the cultivation progress of new species of transgenic fish.

The present invention relates to transgenic pink ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing.

The present invention relates to transgenic purple ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing.

Methods of disrupting germ cell migration and development in a fish embryo by inducing targeted expression of Sdf-1 a or Lif and disruption of the Sdf-1 a gradient or a Lif signaling pathway in the fish embryo have been developed. Plasmid constructs containing a gene encoding Sdf-1 a or Lif and a gene encoding a signaling sequence for targeted expression of Sdf-1 a or Lif have been generated. The plasmids will be administered to a fish or a population of fish to reproductively sterilize the population with efficacy of up to 100%. Transgenic fish of this invention are reproductively incompetent of genetically contaminating a wild fish population.

The present invention relates to transgenic orange ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing.

The present invention relates to transgenic pink ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing.