Preparation of Escherichia coli cell extract for highly productive cell-free protein expression
| Autor: | Rie Nakajima, Takayoshi Matsuda, Takanori Kigawa, Natsuko Matsuda, Shigeyuki Yokoyama, Akiko Tanaka, Takashi Yabuki |
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| Rok vydání: | 2004 |
| Předmět: |
Proteomics
Magnetic Resonance Spectroscopy Cell DNA Recombinant Gene Expression Genomics Biology medicine.disease_cause Cell Fractionation Biochemistry Polymerase Chain Reaction Structural genomics Viral Proteins Structural Biology Gene expression Genetics medicine Protein biosynthesis Escherichia coli Cell-free protein synthesis General Medicine DNA-Directed RNA Polymerases Recombinant Proteins medicine.anatomical_structure |
| Zdroj: | Journal of structural and functional genomics. 5(1-2) |
| ISSN: | 1345-711X |
| Popis: | As structural genomics and proteomics research has become popular, the importance of cell-free protein synthesis systems has been realized for high-throughput expression. Our group has established a high-throughput pipeline for protein sample preparation for structural genomics and proteomics by using cell-free protein synthesis. Among the many procedures for cell-free protein synthesis, the preparation of the cell extract is a crucial step to establish a highly efficient and reproducible workflow. In this article, we describe a detailed protocol for E. coli cell extract preparation for cell-free protein synthesis, which we have developed and routinely use. The cell extract prepared according to this protocol is used for many of our cell-free synthesis applications, including high-throughput protein expression using PCR-amplified templates and large-scale protein production for structure determinations. |
| Databáze: | OpenAIRE |
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