論文 - 詳細
| RRC ID | 60583 |
|---|---|
| 著者 | Zhang JJ, Yamanaka K, Tang X, Moore BS. |
| タイトル | Direct cloning and heterologous expression of natural product biosynthetic gene clusters by transformation-associated recombination. |
| ジャーナル | Methods Enzymol |
| Abstract |
Heterologous expression of natural product biosynthetic gene clusters (BGCs) is a robust approach not only to decipher biosynthetic logic behind natural product (NP) biosynthesis, but also to discover new chemicals from uncharacterized BGCs. This approach largely relies on techniques used for cloning large BGCs into suitable expression vectors. Recently, several whole-pathway direct cloning approaches, including full-length RecE-mediated recombination in Escherichia coli, Cas9-assisted in vitro assembly, and transformation-associated recombination (TAR) in Saccharomyces cerevisiae, have been developed to accelerate BGC isolation. In this chapter, we summarize a protocol for TAR cloning large NP BGCs, detailing the process of choosing TAR plasmids, designing pathway-specific TAR vectors, generating yeast spheroplasts, performing yeast transformation, and heterologously expressing BGCs in various host strains. We believe that the established platforms can accelerate the process of discovering new NPs, understanding NP biosynthetic logic, and engineering biosynthetic pathways. |
| 巻・号 | 621 |
| ページ | 87-110 |
| 公開日 | 2019-1-1 |
| DOI | 10.1016/bs.mie.2019.02.026 |
| PII | S0076-6879(19)30045-X |
| PMID | 31128791 |
| PMC | PMC6555405 |
| MeSH | Bacteria / genetics* Bacteria / metabolism Biological Products / metabolism Biosynthetic Pathways Cloning, Molecular / methods* Firmicutes / genetics Firmicutes / metabolism Genetic Engineering / methods Micromonosporaceae / genetics Micromonosporaceae / metabolism Multigene Family* Proteobacteria / genetics Proteobacteria / metabolism Recombination, Genetic* Saccharomyces cerevisiae / genetics* Saccharomyces cerevisiae / metabolism Streptomyces / genetics Streptomyces / metabolism |
| IF | 1.862 |
| 引用数 | 7 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 6 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 一般微生物 | JCM20779 |