Gateway-compatible lentiviral transfer vectors for ubiquitin promoter driven expression of fluorescent fusion proteins

Niklas Krupka, Padraig Strappe, Jurgen Götz, Lars M. Ittner

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Lentiviral gene delivery has become widely used. Similarly, the Gateway cloning technology that allows restriction-independent cloning of genes into target vectors is becoming increasingly popular. Here, we have generated two Gateway-compatible lentiviral transfer vectors for expression of carboxy-terminal fluorescence tagged fusion proteins, pLVU/GFP and pLVU/RED. We used a restriction enzyme-independent PCR-based approach to introduce the carboxy-terminal fluorescence tags, EmGFP and DsRed, respectively. Both vectors combine the advantages of restriction enzyme/ligation-independent cloning using the Gateway system with a attR1-CmR-ccdB-attR2 recombination cassette, together with expression of fluorescence tagged fusion proteins driven by the robust mammalian ubiquitin C (UbC) promoter. We tested the vectors by expressing different proteins together with the carboxy-terminal fluorescence tags in 293T and SH-SY5Y cells. Both pLVU/GFP and pLVU/RED can be utilized in different experiments, including protein localization studies and live-cell in vivo imaging.
Original languageEnglish
Pages (from-to)155-160
Number of pages6
JournalPlasmid
Volume63
Issue number3
DOIs
Publication statusPublished - 2010

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