相关实验视频
Updated: May 20, 2025

10:09
Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX
Published on: June 27, 2017
13.2K
外星染色体作为一个新的平台,用于Kluyveromyces marxianus中的多个基因表达
Yilin Lyu1,2, Jungang Zhou1,2, Yao Yu1,2
1State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200438, China.
Microorganisms
|March 27, 2025
概括
外来染色体为Kluyveromyces marxianus酵母中的多个基因表达提供了一个稳定的平台. 这一突破可以通过代谢工程和合成生物学来提高有价值产品的生产.
科学领域:
- 微生物学 微生物学
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
背景情况:
- Kluyveromyces marxianus 是一个有前途的酵母宿主,可以生产各种产品.
- 在K. marxianus中表达多个基因存在挑战,因为对cis-regulatory元素和插入位的知识有限.
- 之前的一项研究成功地将Saccharomyces cerevisiae染色体I (R1) 转移到K. marxianus中,创造了KS-R1杂交酵母.
研究的目的:
- 研究外来R1染色体作为K. marxianus.多基因整合和表达的稳定平台的实用性.
- 在R1染色体上识别和描述稳定的转录位点.
- 证明R1在增强异质基因表达和产物形成方面的潜力.
主要方法:
- 通过删除必需的K. marxianus基因并将它们补充为R1 orthologs来实现R1的稳定传播.
- 在不同的条件下,R1上的各种位点具有稳定的转录活性.
- 在R1的六个位置插入绿色光蛋白 (GFP),以评估表达水平.
- 五个R1基因被替换为来自Pichia pastoris的二硫化键形成基因,以评估异质基因表达和酶生产.
主要成果:
- 在没有选择性压力的情况下,异种R1染色体的稳定传播得到了实现.
- 在R1上的几个位点在各种条件下表现出稳定的转录活性.
- 在六个测试的位点中,GFP表达有所不同,其中两个位点表达明显高于单位集成.
- 在不同的R1位点共同表达了五个P. pastoris二硫化键形成基因,导致了活跃的基因表达和增强的BadGLA和TeGlaA葡萄糖胺酶的产生.
结论:
- 外来染色体作为一个稳定和多功能平台,用于协调K. marxianus.多基因表达.
- 这种方法为推进代谢工程和合成生物学应用提供了有价值的工具.
- R1染色体提供了一个强大的系统,用于增强酵母中异质产品的生产.
相关概念视频
DNA Microarrays
17.1K
Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
17.1K
Reporter Genes
11.1K
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
11.1K
Exon Recombination
3.5K
The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes.
Exon shuffling follows “splice frame rules.” Each exon...
Exon shuffling follows “splice frame rules.” Each exon...
3.5K
Chromatin Position Affects Gene Expression
23.2K
Chromatin is the massive complex of DNA and proteins packaged inside the nucleus. The complexity of chromatin folding and how it is packaged inside the nucleus greatly influences access to genetic information. Generally, the nucleus' periphery is considered transcriptionally repressive, while the cell's interior is considered a transcriptionally active area.
Topologically Associated Domains (TADs)
The 3-dimensional positioning of chromatin in the nucleus influences the...
Topologically Associated Domains (TADs)
The 3-dimensional positioning of chromatin in the nucleus influences the...
23.2K
Genome Size and the Evolution of New Genes
7.8K
While every living organism has a genome of some kind (be it RNA, or DNA), there is considerable variation in the sizes of these blueprints. One major factor that impacts genome size is whether the organism is prokaryotic or eukaryotic. In prokaryotes, the genome contains little to no non-coding sequence, such that genes are tightly clustered in groups or operons sequentially along the chromosome. Conversely, the genes in eukaryotes are punctuated by long stretches of non-coding sequence.
7.8K
Cell Specific Gene Expression
13.3K
Multicellular organisms contain a variety of structurally and functionally distinct cell types, but the DNA in all the cells originated from the same parent cells. The differences in the cells can be attributed to the differential gene expression. Liver cells, whose functions include detoxification of blood, production of bile to metabolize fats, and synthesis of proteins essential for metabolism, must express a specific set of genes to perform their functions. Gene expression also varies with...
13.3K

