在哺乳动物细胞中,细菌病毒和等离子体基因传递的全面比较
Maria Toth1, Manuel Reithofer1, Gregory Dutra2
1Department of Biotechnology, University of Natural Resources and Life Sciences Vienna, 1190 Vienna, AT, Austria.
Viruses
|March 28, 2024
概括
使用baculovirus的BacMam平台在哺乳动物细胞中提供了优越的重组蛋白质生产,与标准转染方法相比. 这种baculovirus系统展示了高效的短暂和稳定的表达,提高了产量和可扩展性.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 哺乳动物细胞蛋白质的生产依赖于短暂的转染或稳定的细胞系的产生,两者都有局限性.
- 基于baculovirus的转导系统BacMam提供了一个用于重组蛋白质表达的替代方案.
研究的目的:
- 为了比较BacMam转导与六种转染剂的短暂和稳定的蛋白质表达.
- 为了优化BacMam过程,包括表达增强和baculovirus净化.
- 为了评估HEK293-6E,CHO-K1和Vero细胞系的BacMam效率.
主要方法:
- 对六种转染剂与baculovirus转导的比较分析.
- 评估短暂和稳定的蛋白质表达 (ACE2-eGFP模型).
- 过程优化包括酸盐,TSA和baculovirus超离心.
主要成果:
- 在所有测试的细胞系中,BacMam转导在效率方面显著超过了所有转染剂.
- 使用甲酸盐增强剂实现了蛋白质表达的18倍增加.
- 超离心提高了baculovirus转导效率和蛋白质产量,证明了扩大规模的可行性.
结论:
- 在哺乳动物重组蛋白质生产中,BacMam平台优于标准转染.
- 细菌病毒高效地转化多种细胞系,以实现暂时和稳定的表达.
- BacMam为复合蛋白制造提供了一个可扩展和高效的解决方案.
相关概念视频
01:10Intracellular Movement of Viruses and Bacteria
Intracellular bacteria and viruses often comprise a group of highly infectious pathogens that can cause several diseases. Bacterial pathogens include those belonging to the genus Rickettsia responsible for conditions such as rocky mountain spotted fever and the Mediterranean spotted fever; Chlamydia, a genus responsible for a sexually transmitted disease; Coxiella burnetii, an agent responsible for Q fever. Viral pathogens include vaccinia—a poxvirus, and herpes simplex virus—a virus that...
01:19Conjugation
Conjugation is a form of horizontal gene transfer that primarily occurs in bacteria and some archaea, promoting genetic diversity and adaptation. Bacteria can acquire resistance genes through conjugative plasmids, allowing them to survive antibiotic treatments that would otherwise be lethal. This process involves direct contact between cells through specialized structures such as the sex pilus and is mediated by conjugative plasmids, including the F (fertility) factor.Conjugation requires...
01:16Transduction
Among the three main modes of HGT—transformation, conjugation, and transduction—transduction is unique in that it is mediated by bacteriophages, or bacterial viruses.Transduction occurs in two ways. Generalized transduction occurs during the lytic cycle of a bacteriophage infection. In this process, bacteriophages infect bacterial cells, replicate within them, and ultimately cause cell lysis, releasing newly assembled virions. Occasionally, random fragments of the bacterial genome are...
01:29Modern Molecular Taxonomy
Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...
01:08Evolution of Microbial Genome
Microbial genome evolution is a highly dynamic process shaped by continual gene gain and loss across species and strains. This genomic flexibility allows microorganisms to adapt rapidly to environmental pressures and interactions with other organisms. Central to understanding this diversity is the distinction between the core and pan genomes.The core genome comprises the genes shared by all sampled strains of a species, representing essential functions needed for fundamental cellular processes.
01:18Bacterial Gastroenteritis
Bacterial gastroenteritis, characterized by diarrhea, abdominal cramps, and vomiting, is often caused by ingestion of contaminated food or water and is frequently associated with pathogenic Escherichia coli strains. These microbes exploit two principal mechanisms to inflict disease.Shiga toxin–producing E. coli, also referred to as STEC—notably O157:H7—release Shiga toxins that target ribosomes, blocking protein synthesis. The B subunit of the toxin binds the host glycolipid receptor...