Data/hora: 16/06/2024 04:59:08

Provedor de dados: | 17
País: | United Kingdom
Título: | Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interactions in living cells
Autores: | Jian-Ping Lu; Laura Beatty; Jehonathan  Pinthus.
Data: | 2008-01-31
Ano: | 2008
Palavras-chave: | Biotechnology; Molecular Cell Biology.
Resumo: | Identification of novel protein interactions and their mediators is fundamental in understanding cellular processes and is necessary for protein-targeted therapy. Evidently high throughput formatting of these applications in living cells would be beneficial, however no adequate system exists. We present a novel platform technology for the high throughput screening and verification of protein interactions in living cells. The platform&amp;#x2019;s series of Dual Expression Recombinase Based (DERB) destiny vectors individually encode two sets of recombinase recognizable sequences for inserting the protein open reading frame (ORF) of interest, two sets of promoters and reporter tags in frame with the ORFs for detecting interactions. Introduction into living cells (prokaryotic and eukaryotic) enables the detection of protein interactions by fluorescence resonance energy transfer (FRET) or bimolecular fluorescence complementation (BiFC). The DERB platform shows advantages over current commercialized systems by introducing recombinase based cloning and compatible accepting vectors validated through proof-of-principle experiments and the identification of an unknown interaction.
Tipo: | Manuscript

Identificador:

http://precedings.nature.com/documents/1550/version/2

oai:nature.com:10101/npre.2008.1550.2

http://hdl.handle.net/10101/npre.2008.1550.2

Fonte: | Nature Precedings
Fonte: | Creative Commons Attribution 3.0 License