Construction of Transgenic Goldfish Based on Tol2 Transposable Element

Abstract

Abstract: Medaka Tol2 transposon(Transposable element of Oryzias latipes, number 2)was the first natural active transposon in vertebrate, which had been successfully applied in transgenic research of variety model animals. The sequence containing five exons of grass carp growth hormone(GH), carp beta-actin promoter and multiple restriction enzyme cutting sites was amplified based on pCAgcGH plasmid, and was inserted into pTol2-MCS-EGFP plasmid to construct recombinant pTol2-GH-EGFP plasmid. The recombinant pTol2-GH-EGFP plasmid and the transposase mRNA synthesized in vitro were coinjected into fertilized eggs of goldfish by microinjection. The transgenic goldfish were selected by observing green fluorescence and PCR, and expression rate was 17.3%. The construction of transgenic goldfish based on Tol2 transposable element, not only expanded the range of Tol2 transposon application, but also would be useful for studies of transgene and gene expression in ornamental fish.

Key words: Tol2 transposon, Transgenesis, Goldfish

Dong Ying, Hu Hongxia, Wang Wei, Tian Zhaohui. Construction of Transgenic Goldfish Based on Tol2 Transposable Element[J]. Biotechnology Bulletin, 2014, 0(5): 122-128.

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