使用基因密码扩展编制的无珀病毒的HIV-1信封的点击标签协议
Wang Xu1, Narendra Kumar Gonepudi1, Maolin Lu1
1Department of Cellular and Molecular Biology, School of Medicine, University of Texas at Tyler Health Science Center, Tyler, TX 75708, USA.
STAR protocols
|July 27, 2025
概括
这项研究详细介绍了一种用于在病毒颗粒上光标记HIV-1包膜糖蛋白的新方法. 该技术允许在感染期间直接可视化病毒蛋白质动态.
科学领域:
- 生物化学 生物化学
- 病毒学 病毒学
- 分子生物学分子生物学
背景情况:
- 对病毒蛋白动态的直接可视化对于理解HIV-1感染至关重要.
- 目前用于标记病毒蛋白的现有方法在特异性和应用方面存在局限性.
研究的目的:
- 关于在病毒体上对HIV-1包裹 (Env) 糖蛋白进行特定地点的光标记的协议.
- 为了使HIV-1蛋白质动态和在感染生命周期中的贩运能够直接可视化.
主要方法:
- 利用遗传密码扩展和点击化学进行标记.
- 使用HEK293T细胞产生无珀的HIV-1与非正规氨基酸 (ncAA).
- 结合光体与病毒包膜糖蛋白上结合的ncAA.
主要成果:
- 在病毒体上成功地实现了HIV-1 Env糖蛋白的特定位点光标签.
- 该协议有助于通过感染周期跟踪病毒蛋白质.
结论:
- 本协议为研究HIV-1蛋白质动态提供了一种实用方法.
- 该方法结合了珀抑制和点击化学,用于有效的病毒蛋白跟踪.
相关概念视频
Genetic Lingo
Overview
Viral Recombination
Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.
Viral Mutations
A mutation is a change in the sequence of bases of DNA or RNA in a genome. Some mutations occur during replication of the genome due to errors made by the polymerase enzymes that replicate DNA or RNA. Unlike DNA polymerase, RNA polymerase is prone to errors because it is not capable of “proofreading” its work. Viruses with RNA-based genomes, like HIV, therefore accrue mutations faster than viruses with DNA-based genomes. Because mutation and recombination provide the raw material for adaptive...
Retroviruses
Retroviruses and retrotransposons both insert copies of their genetic elements into the genome of the host cell. Thus, the viral genes are passed on when the host genome is replicated or translated. A typical retroviral DNA sequence contains 3-4 genes that encode the different proteins required for its structural assembly and function as a molecular parasite. This DNA is transcribed into a single mRNA, which is very similar in structure to conventional mRNAs, i.e., it is capped at the 5’...
Mechanisms of Retrovirus-induced Cancers
Retroviruses are RNA viruses that have been shown to cause cancers in diverse species, including chickens, mice, cats, and monkeys. The RNA genomes of these viruses are first reverse-transcribed into single and then double-stranded DNA (dsDNA) copies. This dsDNA called proviral DNA then integrates into the host genome. Subsequently, the host cell transcribes the proviral DNA in concert with the chromosomal DNA. This leads to the production of viral RNA and proteins that assemble at the host...
Hepatitis
Hepatitis is an inflammatory condition of the liver most commonly caused by hepatotropic viruses (A–E), though non-infectious causes such as alcohol and drugs also exist.Hepatitis AHepatitis A virus (HAV) is a non-enveloped RNA virus of the Picornaviridae family. It is primarily transmitted via the fecal-oral route, typically through ingestion of contaminated food or water. After ingestion, HAV enters the bloodstream through the oropharynx or intestinal epithelium and reaches the liver. The...


