Synthesis of functional proteins using Escherichia coli extract entrapped in calcium alginate microbeads |
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Authors: | Kwon Yong-Chan Hahn Geun-Hee Huh Kang Moo Kim Dong-Myung |
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Institution: | Department of Fine Chemical Engineering and Chemistry, Chungnam National University, Daejeon 305-764, Korea. |
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Abstract: | In this report, we describe the construction and analysis of a cell-free protein synthesis system immobilized in calcium alginate microbeads. When incubated in a feeding solution that contained amino acids and other low-molecular-weight substrates, the microbeads transcribed and translated coimmobilized DNA into functional proteins. Protein synthesis continued for more than 15 h with the diffusional supply of substrates and removal of by-products. In addition, functional proteins were generated from PCR-amplified genes as efficiently as from plasmid, suggesting that these cell-like microbeads could be used for functional screening of genomic libraries. |
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Keywords: | Cell-free protein synthesis Calcium-alginate Immobilization Micro reactors Artificial cells |
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