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体外磷脂合成方案，结合脂肪酸合成和无细胞基因表达。

Protocol for in vitro phospholipid synthesis combining fatty acid synthesis and cell-free gene expression.

机构信息

Institute for Extra-cutting-edge Science and Technology Avant-garde Research (X-star), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), Natsushima-cho 2-15, Yokosuka 237-0061, Japan.

Department of Biosciences and Informatics, Keio University, Kohoku-ku, Yokohama 223-8522, Japan.

出版信息

STAR Protoc. 2024 Jun 21;5(2):103051. doi: 10.1016/j.xpro.2024.103051. Epub 2024 May 2.

DOI:10.1016/j.xpro.2024.103051
PMID:38700978
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11078692/
Abstract

Phospholipids are important biomolecules for the study of lipidomics, signal transduction, biodiesel, and synthetic biology; however, it is difficult to synthesize and analyze phospholipids in a defined in vitro condition. Here, we present a protocol for in vitro production and quantification of phospholipids. We describe steps for preparing a cell-free system consisting of fatty acid synthesis and a gene expression system that synthesizes acyltransferases on liposomes. The whole reaction can be completed within a day and the products are quantified by liquid chromatography-mass spectrometry. For complete details on the use and execution of this protocol, please refer to Eto et al..

摘要

磷脂是脂质组学、信号转导、生物柴油和合成生物学研究的重要生物分子；然而，在确定的体外条件下合成和分析磷脂是困难的。在这里，我们提出了一种在体外生产和定量磷脂的方案。我们描述了制备无细胞体系的步骤，该体系包括脂肪酸合成和基因表达系统，该系统在脂质体上合成酰基转移酶。整个反应可以在一天内完成，产物通过液相色谱-质谱法进行定量。有关此方案的使用和执行的完整详细信息，请参阅 Eto 等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/438b4b91822a/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/b30cbd1c273b/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/d60f100ff8b5/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/625d7a3751c8/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/5085c52e603c/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/1529ef69c757/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/438b4b91822a/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/b30cbd1c273b/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/d60f100ff8b5/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/625d7a3751c8/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/5085c52e603c/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/1529ef69c757/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5521/11078692/438b4b91822a/gr5.jpg

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