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在Haloferax volcanii中,用于克隆基因和多色光标记的单个和合表达的载体系统

Solenne Ithurbide^1,2, Roshali T de Silva^1,3, Hannah J Brown¹

¹The Australian Institute for Microbiology and Infection, University of Technology Sydney, Ultimo, NSW, 2007, Australia.

Microbiology (Reading, England)

|May 24, 2024

概括

新的等离子体载体使得模型archaeon Haloferax volcanii中的方便的蛋白质生产和光标记成为可能,从而推进了archaeal细胞生物学和生物技术.

科学领域:

考古细胞生物学考古细胞生物学
分子和细胞生物学分子和细胞生物学
生物技术和合成生物学

背景情况:

考古细胞生物学是一个新兴领域,有可能发现基本的细胞过程和进化见解.
在Haloferax volcanii等模型古生物中开发基因操纵工具对于推进该领域至关重要.
蛋白质融合和亚细胞局部化研究是了解蛋白质功能和细胞组织的关键.

研究的目的:

开发多功能等离子体载体,以便在Haloferax volcanii中方便地克隆和生产蛋白质和融合蛋白质.
为了创建N-或C-终端标记的载体,使用各种编码子优化的光蛋白 (FP) 来进行亚细胞局部化研究.
为了促进两个基因或融合基因的合表达,用于双色成像.

主要方法:

开发具有柔性,刚性或半刚性连接器的新型等离子体载体.
在N端和C端进行标记时,将编码子优化的光蛋白 (GFP,mNeonGreen,mCherry,YPet,mTurquoise2,mScarlet-I) 纳入.
通过选细胞骨蛋白FP融合和双色成像来证明载体效用.

主要成果:

成功创建了用于细胞分裂的功能FtsZ1-FP融合和用于运动和细胞发育的CetZ1-FP融合.
确定连接器类型和FP类型都会影响融合蛋白的功能.
在成功的双色成像中,通过使用双联基因表达,证明了标记蛋白质的成功.

结论:

开发的等离子体载体为推进考古分子和细胞生物学提供了必不可少的工具.
这些工具将促进蛋白质生产,局部化研究和Haloferax volcanii的功能分析.
该系统支持在考古生物技术和合成生物学中开发新应用.

关键词:

这就是CetZZ.FtsZZ 在线观看考古学是指古物质 (archaea) 是指古物质.克隆载体的克隆载体细胞骨架细胞骨架光蛋白是一种光蛋白质.

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