Reference - Detail
| RRC ID | 3316 |
|---|---|
| Author | Kawahashi Y, Doi N, Oishi Y, Tsuda C, Takashima H, Baba T, Mori H, Ito T, Yanagawa H. |
| Title | High-throughput fluorescence labelling of full-length cDNA products based on a reconstituted translation system. |
| Journal | J Biochem |
| Abstract |
Although recent advances in fluorescence-based technologies, such as protein microarrays, have made it possible to analyse more than 10,000 proteins at once, there is a bottleneck in the step of preparation of large numbers of fluorescently labelled proteins for the comprehensive analysis of protein-protein interactions. Here we describe two independent methods for high-throughput fluorescence-labelling of full-length cDNA products at their C-termini using a reconstituted translation system containing fluorescent puromycin. For the first method, release factor-free systems were used. For the second method, stop codons were excluded from cDNAs by using a common mismatch primer in mutagenic PCR. These methods yielded large numbers of labelled proteins from cDNA sets of various organisms, such as mouse, yeast and Escherichia coli. |
| Volume | 141(1) |
| Pages | 19-24 |
| Published | 2007-1-1 |
| DOI | 10.1093/jb/mvm003 |
| PII | mvm003 |
| PMID | 17148548 |
| MeSH | Carbocyanines / chemistry Cell-Free System / metabolism DNA, Complementary / chemistry* Escherichia coli / metabolism Escherichia coli Proteins / metabolism Fluorescent Dyes* / chemistry Multienzyme Complexes / metabolism Proteasome Endopeptidase Complex Protein Biosynthesis* Proteomics / methods Puromycin / analogs & derivatives* Saccharomyces cerevisiae Proteins / metabolism |
| IF | 2.476 |
| Times Cited | 4 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Altmetric score |
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 1 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Prokaryotes E. coli | ASKA library |