Tgf2 Transposon Multiple Cloning Sites Cloning and Expression

Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (2): 124-129.

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Tgf2 Transposon Multiple Cloning Sites Cloning and Expression

Tao Ran^1，2, Chang Yumei², Liang Liqun², Tang Ran², Dou Xinjie^2，3, Wang Nan^2，3, Li Mingyun¹,

（1. College of Ocean，Ningbo University，Ningbo 315211；2.China National & Local United Engineering Laboratory of Freshwater Fish Breeding，Key Laboratory of Freshwater Aquatic Biotechnology and Genetic Breeding，Ministry of Agriculture，Heilongjiang River Fisheries Research Institute，Chinese Academy of Fishery Sciences，Harbin 150070；3. College of Fisheries and Life Science，Shanghai Ocean University，Shanghai 201306）

Received:2013-10-08 Online:2014-02-27 Published:2014-02-27

Abstract

Abstract: The goldfish pTgf2-EF1α-EGFP transposon vector was reconstructed by insertion of artificial synthesized multiple colning sites（MCS）. In order to examine its effectiveness, the recombinant vectors of pTgf2-EF1α-MCS-EGFP and pTgf2-EF1α-MCS-MIPS--EGFP containing the exogenous gene, Myo-inositol-3-phosphate synthase（MIPS）were injected into 1-2 cells of zebrafish embryos. Both fluorescence observation of GFP and PCR detection of exogenous gene sizes verified the effectiveness of the reconstructed vector, in spite of slight differences in fluorescent strength of GFP. Moreover, the reconstructed vector has a higher integration efficiency（31.4%）in the genome of zebrafish based on the results. The successful construction of recombinant expression vector displays goldfish Tgf2 transposon can mediate exogenous gene expression in zebrafish and it would be useful to Tgf2 transposon in gene function research of fish.

Tao Ran, Chang Yumei, Liang Liqun, Tang Ran, Dou Xinjie, Wang Nan, Li Mingyun,. Tgf2 Transposon Multiple Cloning Sites Cloning and Expression[J]. Biotechnology Bulletin, 2014, 0(2): 124-129.

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Tgf2 Transposon Multiple Cloning Sites Cloning and Expression

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