Crassostrea gigas elongation factor-1-alpha (EF-1-alpha) promoter, and recombinant vector and application thereof

A technology of EF-1 and long oyster, which is applied in the fields of genetic engineering and molecular biology, and can solve problems such as unsuccessful development
CN110684769AActive Publication Date: 2020-01-14OCEAN UNIV OF CHINA
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
OCEAN UNIV OF CHINA
Publication Date
2020-01-14

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Abstract

The invention discloses a crassostrea gigas elongation factor-1-alpha (EF-1-alpha) promoter as well as a recombinant vector and application thereof. The invention provides an EF-1-alpha promoter. TheEF-1-alpha promoter is a high-expression housekeeping gene promoter which can be expressed in early stage of embryo development of crassostrea gigas. The EF-1-alpha promoter is specifically expressedas follows: designing a specific primer; amplifying a fragment of the crassostrea gigas EF-1-alpha promoter; inserting the crassostrea gigas EF-1-alpha promoter into a vector containing a GFP reportergene by utilizing In-fusion connection so as to obtain a recombinant vector; and then, introducing the recombinant vector into fertilized eggs of crassostrea gigas, thereby efficiently driving expression of the GFP reporter gene in embryos and larvae of crassostrea gigas. A universal-type efficient promoter sequence for crassostrea gigas is prepared according to the invention; and thus, importanttechnical basis is provided for development of CRISPR / cas9 targeted gene editing technology.
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Description

technical field

[0001] The invention relates to the fields of genetic engineering and molecular biology, in particular to a long oyster EF-1α promoter and its recombinant vector and application. Background technique

[0002] As a worldwide distribution of marine shellfish, long oyster has important economic value. In recent years, my country's long oyster genetics and breeding work has made great progress, and new varieties of long oyster "Haida No. 1" and "Haida No. 2" have been bred successively. The methods mainly used are traditional artificial selection breeding. In recent years, the application of CRISPR / Cas9 targeted gene editing technology in animal and plant genetic improvement and breed breeding has attracted widespread attention. The method of introducing Cas9 protein + sgRNA into fertilized eggs by microinjection can achieve efficient and precise target gene editing in long oyster. However, this method is not suitable for large-scale processing of fertilized eg...

Claims

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