| pubmed-article:10404162 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:10404162 | lifeskim:mentions | umls-concept:C0036576 | lld:lifeskim |
| pubmed-article:10404162 | lifeskim:mentions | umls-concept:C0872079 | lld:lifeskim |
| pubmed-article:10404162 | lifeskim:mentions | umls-concept:C1515655 | lld:lifeskim |
| pubmed-article:10404162 | pubmed:issue | 7 | lld:pubmed |
| pubmed-article:10404162 | pubmed:dateCreated | 1999-9-29 | lld:pubmed |
| pubmed-article:10404162 | pubmed:abstractText | We describe a rapid and efficient in vivo library-versus-library screening strategy for identifying optimally interacting pairs of heterodimerizing polypeptides. Two leucine zipper libraries, semi-randomized at the positions adjacent to the hydrophobic core, were genetically fused to either one of two designed fragments of the enzyme murine dihydrofolate reductase (mDHFR), and cotransformed into Escherichia coli. Interaction between the library polypeptides reconstituted enzymatic activity of mDHFR, allowing bacterial growth. Analysis of the resulting colonies revealed important biases in the zipper sequences relative to the original libraries, which are consistent with selection for stable, heterodimerizing pairs. Using more weakly associating mDHFR fragments, we increased the stringency of selection. We enriched the best-performing leucine zipper pairs by multiple passaging of the pooled, selected colonies in liquid culture, as the best pairs allowed for better bacterial propagation. This competitive growth allowed small differences among the pairs to be amplified, and different sequence positions were enriched at different rates. We applied these selection processes to a library-versus-library sample of 2.0 x 10(6) combinations and selected a novel leucine zipper pair that may be appropriate for use in further in vivo heterodimerization strategies. | lld:pubmed |
| pubmed-article:10404162 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:10404162 | pubmed:language | eng | lld:pubmed |
| pubmed-article:10404162 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:10404162 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:10404162 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
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| pubmed-article:10404162 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:10404162 | pubmed:month | Jul | lld:pubmed |
| pubmed-article:10404162 | pubmed:issn | 1087-0156 | lld:pubmed |
| pubmed-article:10404162 | pubmed:author | pubmed-author:PlückthunAA | lld:pubmed |
| pubmed-article:10404162 | pubmed:author | pubmed-author:ArndtK MKM | lld:pubmed |
| pubmed-article:10404162 | pubmed:author | pubmed-author:MichnickS WSW | lld:pubmed |
| pubmed-article:10404162 | pubmed:author | pubmed-author:PelletierJ... | lld:pubmed |
| pubmed-article:10404162 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:10404162 | pubmed:volume | 17 | lld:pubmed |
| pubmed-article:10404162 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:10404162 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:10404162 | pubmed:pagination | 683-90 | lld:pubmed |
| pubmed-article:10404162 | pubmed:dateRevised | 2006-11-15 | lld:pubmed |
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| pubmed-article:10404162 | pubmed:meshHeading | pubmed-meshheading:10404162... | lld:pubmed |
| pubmed-article:10404162 | pubmed:year | 1999 | lld:pubmed |
| pubmed-article:10404162 | pubmed:articleTitle | An in vivo library-versus-library selection of optimized protein-protein interactions. | lld:pubmed |
| pubmed-article:10404162 | pubmed:affiliation | Département de biochimie, Université de Montréal, Québec, Canada. | lld:pubmed |
| pubmed-article:10404162 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:10404162 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
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