Viral 2A peptides allow expression of multiple proteins from a single ORF in Transgenic zebrafish embryos

被引：145
作者：
Provost, Elayne
Rhee, Jerry
Leach, Steven D.
机构：
[1] Johns Hopkins Univ, Sch Med, Dept Surg, Baltimore, MD 21205 USA
[2] Johns Hopkins Univ, Sch Med, Dept Cell Biol, Baltimore, MD USA
来源：
GENESIS | 2007年 / 45卷 / 10期
关键词：
2A self-cleaving peptide; zebrafish; transgenesis; multicistronic; IRES;
D O I：
10.1002/dvg.20338
中图分类号：
Q [生物科学];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
We have adapted a novel multicistronic gene expression system involving viral peptides to the zebrafish. The viral 2A peptide allows production of multiple protein products from a single transgene. Based on highly inefficient peptide bond formation between glycine and proline residues within the 2A peptide, placement of 2A peptide sequence as a linker region between tandem cDNA?s allows the stoichiometric translation of multiple unfused protein products. To test this system in zebrafish, we generated two different tandem reporter constructs employing eGFP and mCherry, separated by 2A peptide sequence. Using this system, we produced transgenic zebrafish in which fluorophores were produced as independent proteins from a single transcript. The successful application of this technology in zebrafish will be valuable for visually marking transgenic embryos and transgene-expressing cells, or in any situation where reliable expression of multiple transgenes is desired.
引用
收藏
页码：625 / 629
页数：5
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