In vitro selection of GTP-binding proteins by block shuffling of estrogen-receptor fragments |
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Authors: | Toru Tsuji Hiroshi Yanagawa |
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Affiliation: | Department of Biosciences and Informatics, Keio University, Yokohama 223-8522, Japan |
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Abstract: | To what extent has alternative splicing contributed to the evolution of protein-function diversity? We previously constructed a pool of block-deletion mutants of the human estrogen receptor α ligand binding domain by random multi-recombinant PCR. Here we performed iterative in vitro selection of GTP-binding proteins by using the library of mRNA-displayed proteins and GTP-affinity chromatography combined with quantitative real-time PCR. We obtained a novel GTP-binding protein with moderate affinity and substrate-specificity. The results of our in vitro simulation imply that alternative splicing may have contributed substantially to the diversification of protein function during evolution. |
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Keywords: | Alternative splicing Combinatorial protein library In vitro selection mRNA display Protein evolution Synthetic biology |
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