人类口腔上皮细胞作为DNA,RNA和蛋白质分子分析的模型系统
Ajay Kumar Danga1, Pramod C Rath1
1Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India.
Tissue & cell
|March 10, 2024
概括
人类口腔上皮细胞提供了一种简单的方法来分离DNA,RNA和蛋白质. 这些细胞使得分子分析能够用于健康,疾病和人口遗传学研究.
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 基因组学就是基因组学.
背景情况:
- 人类口腔上皮细胞很容易获得.
- 它们为生物样本采集提供了一个非侵入性的来源.
- 以前的分子分析方法通常需要更复杂的样本采集.
研究的目的:
- 建立人类口腔上皮细胞作为分子分析的多功能模型系统.
- 为了证明这些细胞在研究DNA,RNA和蛋白质方面的实用性.
- 验证分析基因表达,蛋白质概况和DNA甲基化方法.
主要方法:
- 从口腔抽样中分离基因组DNA,RNA和蛋白质.
- 聚合酶链反应 (PCR) 和逆转录-PCR (RT-PCR) 用于基因表达分析.
- 西方斑块用于蛋白质表达,SDS-PAGE用于蛋白质分析,MTT测试用于细胞活力.
- 绿色光蛋白 (GFP) 表达和Msp I/Hpa II限制消化用于DNA甲基化分析的过渡性转染.
主要成果:
- 成功分离和分子分析DNA,RNA和蛋白质从口腔细胞.
- 证明了家政基因 (GAPDH,β-actin) 的PCR和RT-PCR.
- 通过西部污染和分析的DNA甲基化模式确认了蛋白质表达.
结论:
- 人口上皮细胞作为全面分子研究的有效和简单的模型.
- 这些方法促进了DNA,RNA和蛋白质的分析,用于各种应用.
- 嘴巴细胞分析支持健康监测,疾病研究,人口遗传学和流行病学.
相关概念视频
00:58Genomic DNA in Eukaryotes
Eukaryotes have large genomes compared to prokaryotes. To fit their genomes into a cell, eukaryotic DNA is packaged extraordinarily tightly inside the nucleus. To achieve this, DNA is tightly wound around proteins called histones, which are packaged into nucleosomes that are joined by linker DNA and coil into chromatin fibers. Additional fibrous proteins further compact the chromatin, which is recognizable as chromosomes during certain phases of cell division.
02:05DNA as a Genetic Template
Two structural features of the DNA molecule provide a basis for the mechanisms of heredity: the four nucleotide bases and its double-stranded nature. The Watson-Crick model of double-helical DNA structure, proposed in 1952, drew heavily upon the X-ray crystallography work of researchers Rosalind Franklin and Maurice Wilkins. Watson, Crick, and Wilkins jointly received the Nobel Prize in Physiology or Medicine for their work in 1962. Franklin was, controversially, excluded from the prize for...
02:21Chromatin Packaging
Each human somatic cell contains 6 billion base-pairs of DNA. Each base-pair is 0.34 nm long, which means that each diploid cell contains a staggering 2 meters of DNA. How is such a long DNA strand packed inside a nucleus measuring only 10 - 20 microns in diameter?
The chromatin
In combination with specialized DNA binding protein called Histones, the DNA double helix forms a compact DNA: protein complex called chromatin. The chromatin itself is further compacted into higher-order structures.
The chromatin
In combination with specialized DNA binding protein called Histones, the DNA double helix forms a compact DNA: protein complex called chromatin. The chromatin itself is further compacted into higher-order structures.
02:05DNA as a Genetic Template
Two structural features of the DNA molecule provide a basis for the mechanisms of heredity: the four nucleotide bases and its double-stranded nature. The Watson-Crick model of double-helical DNA structure, proposed in 1952, drew heavily upon the X-ray crystallography work of researchers Rosalind Franklin and Maurice Wilkins. Watson, Crick, and Wilkins jointly received the Nobel Prize in Physiology or Medicine for their work in 1962. Franklin was, controversially, excluded from the prize for...
01:32Chromatin Packaging
Each human somatic cell contains 6 billion base pairs of DNA. Each base pair is 0.34 nm long, meaning each diploid cell contains a staggering 2 meters of DNA. This long DNA strand is packed inside a nucleus measuring only 10-20 microns in diameter with the help of specialized DNA-binding proteins called histones. Together they form a compact DNA-protein complex called chromatin. The chromatin is further compacted into higher-order structures. The highest level of compaction is achieved during...
01:20Genetic Material
Within the human body, a complex and detailed system of trillions of cells works in unison to sustain life. Each cell houses a nucleus, which contains 46 chromosomes divided into 23 pairs. Chromosomes are highly coiled structures made of the genetic material DNA. These chromosomes are essential carriers of genetic information, with half inherited from the mother through her egg and the other half from the father's sperm, combining to create the unique genetic makeup of an individual.