Generation of high-titer lentivirus for the production of transgenic quail.
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.
Poynter
Greg
G
Division of Biology and Biological Imaging Center, Beckman Institute, California Institute of Technology, Pasadena, CA 91125, USA.
Huss
David
D
Lansford
Rusty
R
D000818:Animals; D030801:Animals, Genetically Modified; D003370:Coturnix; D018014:Gene Transfer Techniques; D005821:Genetic Techniques; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D014461:Ultracentrifugation / Q000379:methods; D014759:Viral Envelope Proteins / Q000378:metabolism
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins