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Parallel gene cloning and protein production in multiple expression systems
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
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
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
Keywords:parallel cloning  Pichia pastoris  baculovirus  inclusion bodies  protease  tag
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