A Yeast BiFC-seq Method for Genome-wide Interactome Mapping |
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Affiliation: | 1. State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing 102206, China;2. Department of Biotechnology, Beijing Institute of Radiation Medicine, Beijing 100850, China;3. Department of Rehabilitation Medicine, Nan Lou; Key Laboratory of Wound Repair and Regeneration of PLA, College of Life Sciences, Chinese PLA General Hospital, Beijing 100853, China;4. School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China |
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Abstract: | Genome-wide physical protein–protein interaction (PPI) mapping remains a major challenge for current technologies. Here, we reported a high-efficiency BiFC-seq method, yeast-enhanced green fluorescent protein-based bimolecular fluorescence complementation (yEGFP-BiFC) coupled with next-generation DNA sequencing, for interactome mapping. We first applied yEGFP-BiFC method to systematically investigate an intraviral network of the Ebola virus. Two-thirds (9/14) of known interactions of EBOV were recaptured, and five novel interactions were discovered. Next, we used the BiFC-seq method to map the interactome of the tumor protein p53. We identified 97 interactors of p53, more than three-quarters of which were novel. Furthermore, in a more complex background, we screened potential interactors by pooling two BiFC libraries together and revealed a network of 229 interactions among 205 proteins. These results show that BiFC-seq is a highly sensitive, rapid, and economical method for genome-wide interactome mapping. |
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Keywords: | Bimolecular fluorescence complementation Protein–protein interaction High-throughput Next-generation sequencing |
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