Poynter Greg, Huss David, Lansford Rusty
Division of Biology and Biological Imaging Center, Beckman Institute, California Institute of Technology, Pasadena, CA 91125, USA.
Cold Spring Harb Protoc. 2009 Jan;2009(1):pdb.prot5117. doi: 10.1101/pdb.prot5117.
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.
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。
