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Compartmentalized linkage of genes encoding interacting protein pairs
Author(s) -
Nirantar Saurabh R.,
Ghadessy Farid J.
Publication year - 2011
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201000643
Subject(s) - interactome , computational biology , biology , proteomics , context (archaeology) , dna sequencing , gene , genomic library , primer (cosmetics) , two hybrid screening , genetics , base sequence , chemistry , paleontology , organic chemistry
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 i CLIP (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 i CLIP library for interactants by “PCR‐pulldown,” using a primer specific to a gene of interest along with a universal primer. i CLIP 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.