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Novel transgenic medaka, gene fragments

A gene fragment, medaka technology, applied in genetic engineering, plant genetic improvement, application, etc., to reduce breeding costs and maintain the effect of transgenic quantity

Inactive Publication Date: 2005-09-07
TAIKONG +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a transgenic green fluorescent medaka fish has been published in the literature, the methods and conditions for different types of transgenic fish to produce other fluorescent genes (such as red fluorescent protein) are not the same, and due to the gene construction and gene expression of transgenic technology, The different strategies and uncertain factors of genetic inheritance make it impossible for us to easily deduce from the existing technology

Method used

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  • Novel transgenic medaka, gene fragments
  • Novel transgenic medaka, gene fragments
  • Novel transgenic medaka, gene fragments

Examples

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Embodiment Construction

[0043] The following examples are non-limiting and are only representative of various possibilities and features in the present invention.

[0044] To generate red fluorescent medaka fish:

[0045] 1. A commercially available plasmid construct, pDsRed2-1 (Clontech), was used to prepare expression vectors.

[0046] 2. The DsRed fragment comes from the pDsRED2-1 plasmid. The CMV promoter and two inverted terminal repeats (ITRs) of AAV were ligated to the DsRed fragment, such as figure 1 The plasmid construct pDsRed2-1-ITR was prepared as described. The pDsRed2-1-ITR plasmid construct has better expression stability.

[0047] 3. Generation of a new plasmid construct: pβ-DsRed2-1-ITR

[0048] Such as figure 1 As shown, the medaka β-actin promoter was obtained by cutting the plasmid construct pOBA-109 with NcoI and EcoRI restriction enzymes. First use NcoI to fill in the ends, and then cut with EcoRI to generate a 4kb fragment.

[0049] Such as figure 1 As shown, the CMV pr...

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Abstract

The invention also relates to a novel nucleic acid fragment including: (1)medaka beta-actin promotor; (2) a fluoyescent gene; and (3) inverted terminal repeats (ITR) of adeno-associated virus. The invention further relates to a plasmid comprising the nucleic acid fragment of the invention.

Description

technical field [0001] The present invention relates to a novel nucleic acid fragment. Background technique [0002] Ornamental fish is part of the fishery industry and has a global market. Therefore, using DNA recombination and transgenic technology to change the phenotype of ornamental fish has a huge market value. [0003] Transgenic fish studies utilize genes driven by heterologous and homologous regulatory elements and produced by compositional or tissue-specific expression of genes. Regulatory elements include antifreeze protein, mouse metallothionein, chicken δ-crystalline, carp β-actin, salmon histone H3 and Carp α-globin (α-globin) gene and so on. However, the use of these DNA elements in transgenic fish has significant disadvantages, including low expression efficiency and mosaic expression of the transgene. [0004] Microinjection of the lac reporter gene driven by the mekada β-actin promoter into medaka eggs results in transient expression of the lacZ gene, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027C07K14/435C12N15/12C12N15/63C12N15/85C12N15/861
CPCA01K2217/05C07K14/43595A01K2267/02A01K2227/40A01K67/0275C12N15/8509
Inventor 蔡怀桢
Owner TAIKONG
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