In vitro selection of proteins with desired characteristics using mRNA-display
نویسندگان
چکیده
منابع مشابه
In vitro selection of Jun-associated proteins using mRNA display.
Although yeast two-hybrid assay and biochemical methods combined with mass spectrometry have been successfully employed for the analyses of protein-protein interactions in the field of proteomics, these methods encounter various difficulties arising from the usage of living cells, including inability to analyze toxic proteins and restriction of testable interaction conditions. Totally in vitro ...
متن کاملmRNA display: diversity matters during in vitro selection.
D Wilson, Anthony Keefe, and Jack Szostak have made a wonderful contribution to the changing world of peptide and protein in vitro evolution and selection. In a previous issue of PNAS (1), they demonstrated that the expected (and observed) weak affinities of peptides (from, for example, bead or phage display) can be enhanced by using ‘‘mRNA display.’’ The present work is a large step forward. T...
متن کاملIn vitro selection of state-specific peptide modulators of G protein signaling using mRNA display.
The G protein regulatory (GPR) motif is a approximately 20-residue conserved domain that acts as a guanine dissociation inhibitor (GDI) for G(i/o)(alpha) subunits. Here, we describe the isolation of peptides derived from a GPR consensus sequence using mRNA display selection libraries. Biotinylated G(i)(alpha)(1), modified at either the N or C terminus, serves as a high-affinity binding target f...
متن کاملIn vitro evolution of single-chain antibodies using mRNA display
Here we describe the application of the in vitro virus mRNA display method, which involves covalent linkage of an in vitro-synthesized antibody (phenotype) to its encoding mRNA (genotype) through puromycin, for in vitro evolution of single-chain Fv (scFv) antibody fragments. To establish the validity of this approach to directed antibody evolution, we used random mutagenesis by error-prone DNA ...
متن کاملAffinity selection of DNA-binding protein complexes using mRNA display
Comprehensive analysis of DNA-protein interactions is important for mapping transcriptional regulatory networks on a genome-wide level. Here we present a new application of mRNA display for in vitro selection of DNA-binding protein heterodimeric complexes. Under improved selection conditions using a TPA-responsive element (TRE) as a bait DNA, known interactors c-fos and c-jun were simultaneousl...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Methods
سال: 2013
ISSN: 1046-2023
DOI: 10.1016/j.ymeth.2012.11.004