Engineering of chimeric eukaryotic/bacterial Rubisco large subunits in Escherichia coli
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- Koay Teng Wei
- Department of Chemical Science, Faculty of Science, Universiti Tunku Abdul Rahman
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- Wong Hann Ling
- Department of Biological Science, Faculty of Science, Universiti Tunku Abdul Rahman
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- Lim Boon Hoe
- Department of Chemical Science, Faculty of Science, Universiti Tunku Abdul Rahman
書誌事項
- タイトル別名
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- Engineering of chimeric eukaryotic/bacterial Rubisco large subunits in <i>Escherichia coli</i>
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説明
<p>Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a rate-limiting photosynthetic enzyme that catalyzes carbon fixation in the Calvin cycle. Much interest has been devoted to engineering this ubiquitous enzyme with the goal of increasing plant growth. However, experiments that have successfully produced improved Rubisco variants, via directed evolution in Escherichia coli, are limited to bacterial Rubisco because the eukaryotic holoenzyme cannot be produced in E. coli. The present study attempts to determine the specific differences between bacterial and eukaryotic Rubisco large subunit primary structure that are responsible for preventing heterologous eukaryotic holoenzyme formation in E. coli. A series of chimeric Synechococcus Rubiscos were created in which different sections of the large subunit were swapped with those of the homologous Chlamydomonas Rubisco. Chimeric holoenzymes that can form in vivo would indicate that differences within the swapped sections do not disrupt holoenzyme formation. Large subunit residues 1–97, 198–247 and 448–472 were successfully swapped without inhibiting holoenzyme formation. In all ten chimeras, protein expression was observed for the separate subunits at a detectable level. As a first approximation, the regions that can tolerate swapping may be targets for future engineering.</p>
収録刊行物
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- Genes & Genetic Systems
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Genes & Genetic Systems 91 (3), 139-150, 2016
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詳細情報 詳細情報について
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- CRID
- 1390001205474519680
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- NII論文ID
- 130005170246
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- NII書誌ID
- AA11077421
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- ISSN
- 18805779
- 13417568
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- NDL書誌ID
- 027722939
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- PubMed
- 27301279
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- 本文言語コード
- en
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- 資料種別
- journal article
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- データソース種別
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- NDLサーチ
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- PubMed
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- 使用不可