PiggyBac转座系统在转mCherry基因斑马鱼上的应用

doi:10.13560/j.cnki.biotech.bull.1985.2016.02.027

摘要/Abstract

摘要： 改造了PiggyBac转座子供体质粒5'-PTK-3'，将红色荧光蛋白基因(mCherry)定向插入其中，构建得到了基因捕获载体质粒：PTK-mCherry。通过显微注射，将PiggyBac 转座子供体质粒与PiggyBac转座酶mRNA共注射于斑马鱼受精卵中，筛选得到了mCherry的表达在时间及空间上均不同的转基因斑马鱼，统计表明mCherry在F0代中有8.2%的表达率，PiggyBac转座子能够在斑马鱼中进行有效转座且捕获到不同基因，有效实现突变体的筛选。

关键词: PiggyBac转座子, 斑马鱼, 基因捕获

Abstract: The red fluorescent protein gene(mCherry)was inserted into the modified PiggyBac transposon donor plasmid 5'-PTK-3', and the vector plasmid for gene trap, PTK-mCherry was constructed.PTK-mCherry and PiggyBac transposase mRNA were co-injected into fertilized eggs by microinjection, and the zebrafishes expressing mCherry in different temporally and spatially patterns were acquired.The statistics revealed that the expression rate of mCherry in F0 was 8.2%, indicating that PiggyBac transposon efficiently translocated and captured the different genes in zebrafish, and the efficient screening of mutants was achieved.

Key words: PiggyBac transposon, zebrafish, gene trap

曹守莹, 白长存. PiggyBac转座系统在转mCherry基因斑马鱼上的应用[J]. 生物技术通报, 2016, 32(2): 185-191.

CAO Shou-ying, BAI Chang-cun. The Application of the PiggyBac Transposon System in mCherry-transgenic Danio rerio[J]. Biotechnology Bulletin, 2016, 32(2): 185-191.

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