基因工程和提高温度增强了与Trichoderma reesei中的cdna1促进体的重组蛋白质生产
Shanshan Jiang1, Yue Wang1, Qin Liu1
1State Key Laboratory of Microbial Technology, Shandong University, 72 Binhai Road, Qingdao, 266237, China.
Bioresources and bioprocessing
|April 22, 2024
概括
在Trichoderma reesei中优化cdna1促进剂显著提高了重组蛋白质的生产. 改进包括促进器工程和热感应,导致β-mannanase产量增加了6.6倍.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 工业微生物学 工业微生物学
背景情况:
- 特里科德尔马·里西 (Trichoderma reesei) 是分泌蛋白质生产的主要宿主.
- cdna1促进体广泛用于构成性蛋白质表达,但需要优化.
- 目前使用cdna1促进剂的生产水平对于工业应用来说是不理想的.
研究的目的:
- 确定cdna1促进体的关键调控区域,以提高蛋白质生产.
- 在各种压力条件下研究cdna1促进体的诱导性.
- 开发策略,以显著提高T. reesei.的异质蛋白产量.
主要方法:
- 通过识别和放大关键的上游区域 (600-700 bp) 来促进工程.
- 增加基因拷贝数以提高转录水平.
- 选最佳的培养温度,以利用促进剂的诱导性 (热感应).
主要成果:
- 在cdna1起始编码子上游的600-700 bp区域被确定为促进器效率的关键.
- 增加该地区的副本数量三倍,异质β-曼酶的产生提高了37.5%.
- cdna1促进体显示出热诱导性,培养在37°C显著增强β-曼酶和多甲氨酸酶的生产,而30°C的培养显著增强β-曼酶和多甲氨酸酶的生产.
结论:
- 这项研究阐明了cdna1促进体的监管要素,促进了其理解.
- 促进器工程,多拷贝基因插入和温度控制的联合策略实现了6.6倍的β-曼酶产量增加.
- 这些发现为增强Trichoderma reesei的重组蛋白质生产提供了有效的方法.
相关概念视频
02:50Transgenic Plants
7.2K
Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...
7.2K
01:09Recombinant DNA
94.3K
Overview
94.3K
01:20The Central Dogma
21.7K
The central dogma explains the flow of genetic information from DNA nucleotides to the amino acid sequence of proteins.
RNA is the Missing Link Between DNA and Proteins
In the early 1900s, scientists discovered that DNA stores all the information needed for cellular functions and that proteins perform most of these functions. However, the mechanisms of converting genetic information into functional proteins remained unknown for many years. Initially, it was believed that a single gene is...
RNA is the Missing Link Between DNA and Proteins
In the early 1900s, scientists discovered that DNA stores all the information needed for cellular functions and that proteins perform most of these functions. However, the mechanisms of converting genetic information into functional proteins remained unknown for many years. Initially, it was believed that a single gene is...
21.7K
00:53Transgenic Organisms
31.1K
Overview
31.1K
02:53Conservative Site-specific Recombination and Phase Variation
6.0K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
6.0K