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

Krupka, Niklas, Strappe, Padraig, Gotz, Jürgen and Ittner, Lars M. (2010) Gateway-compatible lentiviral transfer vectors for ubiquitin promoter driven expression of fluorescent fusion proteins. Plasmid, 63 3: 155-160. doi:10.1016/j.plasmid.2010.01.002


Author Krupka, Niklas
Strappe, Padraig
Gotz, Jürgen
Ittner, Lars M.
Title Gateway-compatible lentiviral transfer vectors for ubiquitin promoter driven expression of fluorescent fusion proteins
Journal name Plasmid   Check publisher's open access policy
ISSN 0147-619X
1095-9890
Publication date 2010-01-01
Year available 2010
Sub-type Article (original research)
DOI 10.1016/j.plasmid.2010.01.002
Open Access Status Not Open Access
Volume 63
Issue 3
Start page 155
End page 160
Total pages 6
Place of publication Maryland Heights, MO, United States
Publisher Academic Press
Language eng
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.
Keyword DsRed
Gateway cloning
GFP
Lentiviral vector
Ligation-independent cloning
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Queensland Brain Institute Publications
 
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