title: Compartmentalized linkage of genes encoding interacting protein pairs.
abstract: Emulsion technology has been successfully applied to the fields of next-generation high-throughput sequencing, protein engineering and clinical diagnostics. Here, we extend its scope to proteomics research by developing and characterizing a method, termed iCLIP (in vitro compartmentalized linkage of interacting partners), which enables genes encoding interacting protein pairs to be linked in a single segment of DNA. This will facilitate archiving of the interactomes from library versus library two-hybrid screens as libraries of linked DNAs. We further demonstrate the ability to interrogate a model yeast two-hybrid iCLIP library for interactants by "PCR-pulldown," using a primer specific to a gene of interest along with a universal primer. iCLIP libraries may also be subjected to high-throughput sequencing to generate interactome information. The applicability of the technique is also demonstrated in the related context of the bacterial two-hybrid system.
authors.affiliation: p53 Laboratory, Immunos, Singapore.
authors.forename: Saurabh R
authors.initials: SR
authors.lastname: Nirantar
authors.forename: Farid J
authors.initials: FJ
authors.lastname: Ghadessy
mesh_terms: D017931:DNA Primers / Q000235:genetics / Q000378:metabolism; D004655:Emulsions / Q000737:chemistry; D015723:Gene Library; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D017422:Sequence Analysis, DNA; D020798:Two-Hybrid System Techniques
chemical_list: D017931:DNA Primers; D004655:Emulsions; D011506:Proteins