优化哺乳动物表达载体的cis-regulatory元素组合
Lu-Yu Zhou1,2,3, Shuang Zhang1,2,3, Li-Yun Li1,2,3
1College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, Henan, People's Republic of China.
Molecular genetics and genomics : MGG
|June 15, 2023
概括
在表达载体中优化 cis-regulatory 元素可显著增强哺乳动物细胞中的基因和重组蛋白质生产. 战略组合,而不仅仅是更多的元素,改善生物合成和诊断的表达.
科学领域:
- 分子生物学分子生物学
- 基因表达规范 基因表达规范
- 生物技术是生物技术.
背景情况:
- 哺乳动物细胞中使用结合 cis 调节元件的基因表达调节尚未得到充分研究.
- 了解这些组合对于优化基因表达载体至关重要.
研究的目的:
- 研究各种ciss-regulatory元素组合对基因表达的影响.
- 优化表达载体以增强蛋白质生产,包括SARS-CoV-2 RBD.
主要方法:
- 用各种组合的促进子,增强子,内子和终止子构建表达向量.
- 使用光显微镜,定量实时PCR (qRT-PCR) 和西部斑点的表达水平的比较.
- 增强绿色光蛋白 (eGFP) 和SARS-CoV-2受体结合域 (RBD) 蛋白的表达分析.
主要成果:
- 优化 cis 作用元素的组合显著调节蛋白质表达水平.
- 一个特定的载体 (CMV增强剂,EF-1α核心促进剂,TEF终止剂) 增加了eGFP表达的3倍和重组RBD蛋白的2.63倍.
- 在结合多个监管元素时,并不总是观察到协同效应.
结论:
- 战略选择和ciss-regulatory元素的组合是最大化基因表达的关键.
- 研究结果为优化生物合成,诊断和治疗中的表达载体提供了见解,包括COVID-19相关的试剂.
- 这项研究为设计高效的基因表达系统提供了一个框架.
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