摘要
改造了Piggy Bac转座子供体质粒5'-PTK-3',将红色荧光蛋白基因(m Cherry)定向插入其中,构建得到了基因捕获载体质粒:PTK-m Cherry。通过显微注射,将Piggy Bac转座子供体质粒与Piggy Bac转座酶m RNA共注射于斑马鱼受精卵中,筛选得到了m Cherry的表达在时间及空间上均不同的转基因斑马鱼,统计表明m Cherry在F0代中有8.2%的表达率,Piggy Bac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。
The red fluorescent protein gene（m Cherry）was inserted into the modified Piggy Bac transposon donor plasmid 5＇-PTK-3＇, and the vector plasmid for gene trap, PTK-m Cherry was constructed. PTK-m Cherry and Piggy Bac transposase m RNA were co-injected into fertilized eggs by microinjection, and the zebrafishes expressing m Cherry in different temporally and spatially patterns were acquired. The statistics revealed that the expression rate of m Cherry in F0 was 8.2%, indicating that Piggy Bac transposon efficiently translocated and captured the different genes in zebrafish, and the efficient screening of mutants was achieved.
出处
《生物技术通报》
CAS
CSCD
北大核心
2016年第2期185-191,共7页
Biotechnology Bulletin
基金
安徽省生化药物工程技术研究中心开放课题(BYEC1302)
蚌埠医学院校自然科学项目(Byky1329)
