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Author:
Date:
2015-08-05
[Abstract] This protocol aims to study intercellular transport of mitochondria, dynamic cellular organelles via tunnelling nanotubes (TNT), a cell membrane extension of cytoskeletal elements. The nanotubular bridges or the tunnelling nanotube highways are one of the emerging new cell-to-cell communication systems which mediates exchange of cellular materials, most importantly as in our observation, mitochondria. Mesenchymal stem cells (MSC) have been well studied to be endowed with a highly efficient intercellular mitochondrial donation ability and this property is now proven crucial to its functional role of rescue in cellular therapy.
[摘要] 此协议旨在研究线粒体，动态细胞器通过隧道纳米管（TNT），细胞骨架元素的细胞膜扩展的细胞间运输。 纳米管桥或隧道纳米管高速公路是新兴的新的细胞到细胞通信系统之一，其介导细胞材料的交换，最重要的是在我们的观察中，线粒体。 间充质干细胞（MSC）已经被充分研究以赋予高效的细胞间线粒体捐赠能力，并且这种性质现在被证明对于其在细胞治疗中的挽救的功能作用是至关重要的。
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Author:
Date:
2014-03-20
[Abstract] This protocol is a simple method for evaluating mutation frequency during African swine fever virus (ASFV) replication, although it could be used also for other DNA viruses (poxvirus, herpesvirus, mimivirus, etc) with minor modifications. In the original Carrascosa et al. (1982), the protocol was carried out with two cloned viruses, BA71Vc (a purified clone from BA71V wild type strain) and vΔpolX (lacking the reparative polymerase, pol X, gene), and two different cell types that can be infected by ASFV, Vero cells and swine macrophages. To facilitate the sequence comparison, a genome fragment containing the B646L gene was amplified by PCR and blunt-end cloned. This gene codes for the major capsid protein (p72) and multiple sequences can be found in the database, so the ...
[摘要] 该协议是用于评估非洲猪瘟病毒（ASFV）复制期间的突变频率的简单方法，尽管其也可以用于其它具有微小修改的DNA病毒（痘病毒，疱疹病毒，mimivirus，等）。 在原始Carrascosa等（1982）中，使用两种克隆的病毒BA71Vc（来自BA71V野生型菌株的纯化克隆）和vΔpolX（缺乏修复聚合酶，pol X， 基因）和可被ASFV，Vero细胞和猪巨噬细胞感染的两种不同细胞类型。 为了便于序列比较，通过PCR扩增含有B646L基因的基因组片段，并平端克隆。 该基因编码主要衣壳蛋白（p72），并且可以在数据库中发现多个序列，因此可以将发现的突变与天然基因变异进行比较。 克隆的片段可以直接从细菌菌落或从小量制备纯化的DNA测序。
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