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火球菌，作为一种新型工具，可用于在古菌类脂双层中生产哺乳动物嗅觉受体。

Haloferax volcanii, as a Novel Tool for Producing Mammalian Olfactory Receptors Embedded in Archaeal Lipid Bilayer.

机构信息

Department of Basic Medical Sciences, Neurosciences and Sensory Organs, University of Bari "Aldo Moro", Piazza Giulio Cesare 11, I-70124 Bari, Italy.

Institute for Chemical-Physical Processes, National Research Council (IPCF-CNR), Bari Unit, via Orabona 4, I-70126 Bari, Italy.

出版信息

Life (Basel). 2015 Mar 9;5(1):770-82. doi: 10.3390/life5010770.

DOI:10.3390/life5010770
PMID:25761264
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4390878/
Abstract

The aim of this study was to explore the possibility of using an archaeal microorganism as a host system for expressing mammalian olfactory receptors (ORs). We have selected the archaeon Haloferax volcanii as a cell host system and one of the most extensively investigated OR, namely I7-OR, whose preferred ligands are short-chain aldehydes, such as octanal, heptanal, nonanal. A novel plasmid has been constructed to express the rat I7-OR, fused with a hexahistidine-tag for protein immunodetection. The presence of the recombinant receptor at a membrane level was demonstrated by immunoblot of the membranes isolated from the transgenic archaeal strain. In addition, the lipid composition of archaeonanosomes containing ORs has been characterized in detail by High-Performance Thin-Layer Chromatography (HPTLC) in combination with Matrix-Assisted Laser Desorption Ionization-Time-Of-Flight/Mass Spectrometry (MALDI-TOF/MS) analysis.

摘要

本研究旨在探索将古菌微生物作为哺乳动物嗅觉受体 (OR) 表达的宿主系统的可能性。我们选择了古菌盐沼盐杆菌作为细胞宿主系统，选择了研究最为广泛的 OR 之一 I7-OR，其首选配体是短链醛，如辛醛、庚醛、壬醛。构建了一种新的质粒来表达与六组氨酸标签融合的大鼠 I7-OR，用于蛋白免疫检测。通过免疫印迹法对从转基因古菌菌株分离的膜进行分析，证明了重组受体在膜水平上的存在。此外，还通过高效薄层色谱 (HPTLC) 结合基质辅助激光解吸电离飞行时间/质谱 (MALDI-TOF/MS) 分析详细表征了含有 OR 的古菌纳米囊泡的脂质组成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/fc183352f67f/life-05-00770-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/7bd75b7d6731/life-05-00770-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/86a4dd87d567/life-05-00770-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/ab73ee91fb30/life-05-00770-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/34ebd7cfdbba/life-05-00770-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/fc183352f67f/life-05-00770-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/7bd75b7d6731/life-05-00770-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/86a4dd87d567/life-05-00770-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/ab73ee91fb30/life-05-00770-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/34ebd7cfdbba/life-05-00770-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ad1/4390878/fc183352f67f/life-05-00770-g005.jpg

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