A modified two-step phage display selection for isolation of polycystin-1 ligands.
Funct Integr Genomics
; 1(3): 193-9, 2000 Nov.
Article
em En
| MEDLINE
| ID: mdl-11793237
ABSTRACT
The identification of proteins that interact with polycystin-1, the product of the autosomal dominant polycystic kidney disease gene, is an important step towards understanding the molecular pathogenesis of the disease. We have developed a two-step approach for the efficient identification of potential polycystin-1 ligands using the T7 phage display system. The first enrichment step of 4-5 rounds of biopanning is followed by a second step of reverse protein overlay assay. Thus, the sequencing efforts are minimized to the analysis of only positive rather than randomly chosen clones from the enriched population as in the standard phage display approach. Most importantly, the modified approach immediately provides the confirmation of the specificity of interaction and discriminates between strong and weak interactions. Here we present several potential interactors with distinct regions of polycystin-1, representing high-affinity binding partners.
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Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
Proteínas
/
Bacteriófago T7
/
Biblioteca de Peptídeos
Idioma:
En
Revista:
Funct Integr Genomics
Ano de publicação:
2000
Tipo de documento:
Article