厦门大学学报（自然科学版）2018，Vol.57Issue(1)：44-49,6.DOI:10.6043/j.issn.0438-0479.201702023
文昌鱼中一个2A肽介导的多基因表达载体构建
Construction of a 2A Peptide-linked Multicistronic Expression Vector for Amphioxus
摘要
Abstract
The multicistronic expression vector mediated by 2A peptide (P2A)possesses advantages such as high cleavage efficien-cy,equimolar amounts of upstream and downstream gene expression,so that it has been widely applied to animal transgenic research. However,up to date,no multicistronic expression vector is available for the application in amphioxus,a promising new model animal for evolutionary development study,therefore we constructed a multicistronic expression vector harboring P2A based on pXT7 tran-scription system.After microinj ecting mRNA transcribed in vitro into amphioxus eggs and fertilization,we observed the embryos un-der a laser confocal microscope and analyzed protein expression using Western blot assay.The results showed that the mRNA was translated and the P2A mediated protein was cleaved effectively in amphioxus embryos.Moreover,the eGFP protein was targeted to nucleus and mCherry protein to cell membrane under the role of corresponding signal peptides.The separation efficiency of the two proteins was about 91％.Furthermore,we constructed a heat-shock protein gene promoter (BbHsp70)mediated P2A multicistronic expression vector,and the experiments showed the efficiency of gene expression and protein separation achieved the desired results.关键词
多基因表达载体/2A肽/热激启动子/共表达/文昌鱼Key words
multicistronic expression vector/2A peptide/heat-shock promoter/co-expression/amphioxus分类
生物科学引用本文复制引用
华俊豪,李光,王义权..文昌鱼中一个2A肽介导的多基因表达载体构建[J].厦门大学学报（自然科学版）,2018,57(1):44-49,6.基金项目
国家自然科学基金(31372188,31471986,31672246) （31372188,31471986,31672246）
