Name: dissecting host-virus interaction in lytic replication of a model herpesvirus
Uploaded: 2011-10-07T12:00:00
Description: Watch this Scientific Journal Video about Dissecting Host-virus Interaction in Lytic Replication of a Model Herpesvirus at JoVE.com

作者要感谢詹姆斯博士（张志坚）提供必要的试剂，包括小牛- / -小鼠，并任日（美国加州洛杉矶大学，药理学和分子医学博士陈（UT西南，分子生物学） ）提供的细菌人工染色体的γHV68本研究。

1。滑鼠感染γHV68功能<ol><li>六到八周，性别相匹配的旧窝小鼠（8〜12只/组），用于病毒性感染。让小鼠适应环境装船后四天（96小时）。 </li><li>使用病毒协议的步骤应进行在一个内阁生物安全2级（BSL2）的标准BSL2注意事项。 </li><li>准备病毒悬浮液（40〜1×10 5个噬斑形成单位[PFU]）在30μl的无菌PBS中每只小鼠的γHV68刚刚之前的实验。保持病毒悬挂在冰上。 </li><li>准备氯胺酮/甲苯​​噻嗪...

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在对病毒感染的反应，MAVS依赖先天免疫信号通路的激活，促进了生产的抗病毒炎性细胞因子10-14。作为模型病毒对人类的致癌性卡波济氏肉瘤相关疱疹病毒，EB病毒3,4使用鼠γHV68的，我们发现，γHV68侵占MAVS-IKKβ的途径，通过促 ​​进病毒裂解性复制转录激活5。采用转基因MEFs和技术在分子病毒学，该协议可以有效识别的信令组件的一个特定的途径是至关重要的γHV68裂解性?...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td>脂质体2000 </td><td> Invitrogen公司</td><td> 11668-019 </td></tr><tr><td> ELECTRO-MAX DH10B感受态细胞</td><td> Invitrogen公司</td><td> 18290-015 </td></tr><tr><td>甲基纤维素</td><td>西格玛</td><td> M0512 </td></tr><tr><td> POWERPREP HP质粒小提系统</td><td> OriGene </td><td> NP100004 </td></tr><tr><td> POWERPREP HP质粒Midiprep系统</td><td> OriGene </td><td> NP100006 </td></tr></tbody></table>

dissecting host-virus interaction in lytic replication of a model herpesvirus

在对病毒感染的反应，主机开发各种防御反应，激活先天免疫信号转导途径，导致抗病毒细胞因子的产生1,2等。为了殖民主机，是专病毒逃避宿主抗病毒反应和处理信号转导通路。揭开宿主 - 病毒相互作用，揭示了对抗病毒感染的新的治疗策略的发展。 小鼠γHV68是密切相关的人类致癌卡波济氏肉瘤相关疱疹病毒和Epsten的EB病毒3,4。在实验室老鼠提供了一个听话的小动物模型来研究宿主反应和病毒感染的整个过程，这是不是人类疱疹病毒在体内的 γHV68感染。在这个协议中，我们提出了一个面板的表型特征的方法和分子剖析主机的信号传输元件的在γHV68裂解replic通报BULLETIN无论是在体内和体外 。在γHV68急性感染体内的转基因小鼠品系的可用性允许在审讯主机信号转导通路中的作用。此外，小鼠胚胎成纤维细胞（MEF中），可以使用从这些缺陷的小鼠品系中分离进一步解剖这些分子的作用，在γHV68裂解复制体外 。 使用病毒学和分子生物学分析，我们可以找出宿主 - 病毒相互作用的分子机制，并确定必要病毒裂解复制的宿主和病毒基因。最后，细菌人工染色体（BAC）的系统有利于突变引入到病毒因子（s），专门中断的宿主 - 病毒相互作用。携带这些突变基因重组γHV68可以用来概括MEFs细胞的表型的γHV68裂解性复制缺陷的在关键主机信号组件。此协议提供了一个很好的策略，询问宿主-病原体相互作用， 在体内和体外的多层次的干预。 最近，我们发现，γHV68侵占先天免疫信号转导通路促进病毒裂解性复制。具体而言，γHV68原发感染激酶IKKβ激活免疫激活IKKβ磷酸化的主病毒的转录因子，复制和反式激活（RTA），促进病毒的转录激活。在这样做时，γHV68有效地夫妇的转录激活机体天然免疫激活，从而促进病毒的转录和裂解性复制。该研究提供了一个很好的例子，可以应用到其他病毒询问宿主 - 病毒相互作用。

Watch this Scientific Journal Video about Dissecting Host-virus Interaction in Lytic Replication of a Model Herpesvirus at JoVE.com

Dissecting Host-virus Interaction in Lytic Replication of a Model Herpesvirus

我们描述了一个协议来识别主机的信号分子裂解性复制的模型疱疹病毒，γ疱疹病毒68（γHV68）的关键角色。利用转基因小鼠品系和胚胎成纤维细胞裂解性复制γHV68，该协议允许表型特征和在病毒裂解复制的病毒 - 宿主相互作用的分子审问。

解剖宿主 - 病毒相互作用模型疱疹病毒裂解性复制

In response to viral infection, a host develops various defensive responses, such as activating innate immune signaling pathways that lead to antiviral ...

Research

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