Method for fish gene in site modification breeding

An in situ modification, gene technology, applied in the direction of genetic engineering, other methods of inserting foreign genetic materials, plant genetic improvement, etc., can solve the problems that restrict the development, popularization and application of transgenic fish, and the biological safety of transgenic fish affects the effective application, etc. Achieve the effect of eliminating doubts about the safety of GMOs and avoiding transfer

Inactive Publication Date: 2005-08-03
SUN YAT SEN UNIV +1
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Problems solved by technology

Although breakthroughs have been made in genetic engineering directional breeding technology, the purpose of genetic engineering breeding has not yet been achieved. Issues such as random integration of exogenous genes, uncontrollable expression of exogenous genes, and biological safety of transgenic fish have seriously affected this gene. The effective application of engineering breeding technology in fish breeding has become a bottleneck restricting the further development and application of transgenic fish research
Therefore, the defect of the prior art is: it adopts the random integration of exogenous genes to complete the genetic engineering breeding

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  • Method for fish gene in site modification breeding
  • Method for fish gene in site modification breeding
  • Method for fish gene in site modification breeding

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Embodiment Construction

[0022] Taking zebrafish as an example, in figure 1 , figure 2 , image 3 Among them, 1 represents the zebrafish growth hormone gene (GH) promoter regulatory sequence 5' flanking sequence (as a homologous long arm) fragment, 2 represents the GH promoter regulatory sequence (GH-promoter) fragment, and 3 represents the growth hormone gene (GH) promoter The 3' flanking sequence (as a homologous short arm) fragment of the regulatory sequence, 4 represents the β-actin gene (β-actin) initiation regulatory sequence fragment, 5 represents two Cre / loxP sequence fragments in the same direction, and 6 represents green fluorescence Protein (GFP) positively marks gene fragments, 7 represents red fluorescent protein (RFP) negatively marks gene fragments, 8 represents forward PCR detection primers, 9 represents reverse PCR detection primers, 10 represents homologous short arm 3' flanking sequences Non-target DNA sequence fragments.

[0023] Embodiments of the present invention are describ...

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Abstract

The present invention aims at providing one homologous recombination method for in-situ fish gene modification breeding. The method includes the following steps: 1) separating specific fish gene flanking sequence; promoter regulating sequence DNA segments to constitute homologous recombination vector with actin gene promoter regulating sequence to replace growth hormone gene promoter regulating sequence; 2) microscopic injecting the homologous recombination vector to zebra fish embryo to screen positive homologous recombination replaced fish fry; and 3) analyzing relative gene expression level and genetic phynotype of the endogenous gene modified fish. The present invention is used in breeding fish variety.

Description

technical field [0001] The invention relates to a method for genetic engineering breeding, in particular to a method for in situ modification and breeding of fish genes. Background technique [0002] The process of genetic breeding is a process of genome manipulation. Genetic engineering breeding is based on the understanding of gene function and regulatory mechanism, and the genome is modified and transformed at the molecular level, so as to obtain directional genetically improved genetic varieties and get rid of the barriers of genetic fortresses between species. shackles, from passive use of genetic resources to active transformation and innovation. In 1982, Palmiter et al. transferred the rat growth hormone gene to the fertilized mouse eggs and bred fast-growing "super mice". This breakthrough achievement ushered in a new era of transgenic animal research. In 1985, the Institute of Hydrobiology of the Chinese Academy of Sciences took the lead in...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027C12N15/12C12N15/62C12N15/65C12N15/85C12N15/89C12Q1/68
Inventor 吴玉萍朱作言胡炜徐安龙
Owner SUN YAT SEN UNIV
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