用于生产转基因鹌鹑的高滴度慢病毒的产生。

Generation of high-titer lentivirus for the production of transgenic quail.

作者信息

Poynter Greg, Huss David, Lansford Rusty

机构信息

Division of Biology and Biological Imaging Center, Beckman Institute, California Institute of Technology, Pasadena, CA 91125, USA.

出版信息

Cold Spring Harb Protoc. 2009 Jan;2009(1):pdb.prot5117. doi: 10.1101/pdb.prot5117.

DOI:10.1101/pdb.prot5117

PMID:20147012

Abstract

This protocol describes how to generate high-titer lentivirus for the production of transgenic Japanese quail. The virus is pseudotyped with vesicular stomatitis virus with the envelope G glycoprotein (VSV-g), which gives a broad infectious range and allows concentration of viral supernatants by ultracentrifugation. Using this method, we typically produce titers >1 x 10(8) transforming units (TU)/mL and recommend using a virus with a titer at least this high for in vivo work.

摘要

本方案描述了如何为转基因日本鹌鹑的生产制备高滴度慢病毒。该病毒用带有包膜G糖蛋白（VSV - g）的水泡性口炎病毒进行假型包装，这赋予了广泛的感染范围，并允许通过超速离心浓缩病毒上清液。使用这种方法，我们通常能产生滴度>1×10⁸转化单位（TU）/mL的病毒，并建议在体内实验中使用滴度至少如此高的病毒。