Parallel gene cloning and protein production in multiple expression systems |
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Authors: | Hui‐Min Wang Yan‐Ping Shih Su‐Ming Hu Wen‐Tsung Lo Hui‐Min Lin Shih‐Shiu Ding Hsin‐Chi Liao Po‐Huang Liang |
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Affiliation: | 1. Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan, ROC;2. Core Facility of Recombinant Protein Production, Academia Sinica, Taipei 11529, Taiwan, ROC |
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Abstract: | To quickly find an optimal expression system for recombinant protein production, a set of vectors with the same restriction sites were constructed for parallel cloning of a target gene and recombinant protein production in prokaryotic and eukaryotic expression systems, simultaneously. These vectors include nucleotide sequences encoding protein tags and protease recognition sites for tag removal, followed by the cloning sites 5′‐EcoRI/3′‐XhoI identical in these vectors for ligating with the sticky‐end PCR product of a target gene. Our vectors allow parallel gene cloning and protein production in multiple expression systems with minimal cloning effort. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 |
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Keywords: | parallel cloning Pichia pastoris baculovirus inclusion bodies protease tag |
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