弗拉单核酸基光蛋白的脊柱循环增加光和稳定性
Tingting Lin1, Yuanyuan Ge1, Qing Gao1
1School of Life Science, Anhui Agricultural University, Hefei, Anhui 230036, P.R. China.
Journal of microbiology and biotechnology
|October 4, 2023
概括
从flavin mononucleotide结合域中设计的循环光蛋白显示出增强的光和稳定性. 这些新型报告器为有氧和无氧环境中的应用提供了更好的性能.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 蛋白质工程是指蛋白质的工程.
背景情况:
- 弗拉单核酸结合蛋白表现出光,但由于光和热稳定性较低,其在记者应用中存在局限性.
- 现有的光记者在不同氧气条件和热应力下难以保持性能.
研究的目的:
- 为记者应用程序设计具有增强性能的循环光蛋白.
- 研究蛋白质循环和链条长度对光,稳定性和在不同氧气条件下的性能的影响.
主要方法:
- 将来自Chlamydomonas reinhardtii的编码子优化的光蛋白纳入使用两个不同的链接器的氧化还原反应分裂蛋白结构.
- 在大肠杆菌SHuffle T7菌株中过度表达前体蛋白质.
- 在试验室中使用减少剂对蛋白进行循环.
主要成果:
- 与线性蛋白相比,带有短链接器的循环蛋白显示出增强的光,增加的热稳定性和对外化酶的抗性.
- 使用长链体 (包括myc-tag和蛋白质酶分裂部位) 的循环处理,与线性化蛋白质相比,导致光度略有增加.
- 有光的E.E. 在缺乏氧气的条件下,过度表达循环记者的大肠杆菌细胞比具有线性结构的细胞更亮.
结论:
- 蛋白质循环显著增强了光和稳定性的黄单核酸结合的基于域的记者.
- 工程循环报告员表现出更好的性能,特别是在无氧条件下.
- 这种循环记者对热友性厌氧生物和其他苛刻的环境中的应用具有前途.
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