相关实验视频

Updated: May 24, 2025

Electron Cryotomography of Bacterial Cells

Electron Cryotomography of Bacterial Cells

Published on: May 6, 2010

通过冷ET在完整组织中的细胞内结构和拷贝逆转移体的快照

Sven Klumpe¹, Kirsten A Senti², Florian Beck¹

¹Research Group CryoEM Technology, Max Planck Institute of Biochemistry, Martinsried, Germany.

|March 6, 2025

概括

长端重复逆转移体 (LTRs) 在Drosophila中形成病毒样粒子 (VLPs). 低温电子断层扫描显示了副本VLP结构和核进口大小选择,在缺乏piRNA的中观察到核转移.

科学领域:

结构生物学
分子生物学
遗传学

背景情况:

长终端重复 (LTR) 反转体是通过整合到宿主基因组进行复制的移动遗传元素.
果Drosophila melanogaster是研究可转移元素的关键模型生物.
在Drosophila中Ty1-copy家族的LTR反转子形成病毒样粒子 (VLPs) 并与逆转病毒相关.

研究的目的:

在Drosophila melanogaster的卵巢细胞和卵室内可视化复制病毒样颗粒 (VLPs).
为了确定副本VLP体的高分辨率结构.
研究核进口和PIWI相互作用RNA (piRNA) 途径在复制VLP定位和复制中的作用.

主要方法:

使用冷聚焦离子束 (FIB) 研磨和提升技术来准备样品.
使用冷电子断层扫描 (cryo-ET) 可视化卵巢细胞和卵室中的副本VLP.
在精子生成过程中对野生类型和TE抑制piRNA通路缺陷的Drosophila进行复制VLP分布的分析.

主要成果:

副本VLP的现场结构得到了7.7 Å的分辨率.
细胞质副本VLP表现出大小异质性,而核VLP是均的,形成密集的.
核进口似乎是复制VLP的尺寸选择器.
在缺乏piRNA通路的中观察到副本转移到核中.

结论:

这项研究提供了高分辨率的结构洞察力,以了解其本地细胞环境中的仿真病毒样粒子 (VLP).
这些发现表明,对于核进口复制VLP的尺寸有选择性机制.
在精子生成过程中,PIWI相互作用RNA (piRNA) 途径在调节拷贝逆转移子核定位方面发挥作用.

关键词:

类植物在LTR反转换中类动物复制化ET 化FIB 在现场结构生物学提升出来的皮RNA路径复制周期

更多相关视频

Cryo-Electron Tomography Remote Data Collection and Subtomogram Averaging

Cryo-Electron Tomography Remote Data Collection and Subtomogram Averaging

Published on: July 12, 2022

Visualization of Organelles In Situ by Cryo-STEM Tomography

Visualization of Organelles In Situ by Cryo-STEM Tomography

Published on: June 23, 2023

相关概念视频

Electron Microscope Tomography and Single-particle Reconstruction

Electron Microscope Tomography and Single-particle Reconstruction

Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...

Cryo-electron Microscopy

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...