id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-008333-1wepke2o	Weisz, Ora A.	Chapter 7 Use of Recombinant Vaccinia Virus Vectors for Cell Biology	2008-02-28		.txt	text/plain	7465	461	56	The recombinant virus is selected, expanded, and used to infect cells, which then express high levels of the foreign protein. Transfection of cells immediately after infection with a vector containing the gene of interest cloned behind a T7 promoter results in rapid and efficient expression of the encoded protein. The use of an early vaccinia promoter to drive the foreign gene is essential for cell biology applications, since the protein will be expressed before most of the cytopathic effects of the virus infection become evident. Before growing a large-scale preparation of the recombinant, we also test that the expressed protein is the correct size by immunoprecipitation from radiolabeled infected cells, followed by electrophoresis in SDS polyacrylamide gels (see Section IV,C,l). Expression is mediated by co-infecting the cells with another vaccinia virus encoding T7 RNA polymerase (vTF7-3). This chapter has discussed the preparation and use of recombinant vaccinia viruses to express proteins in mammalian cells.	./cache/cord-008333-1wepke2o.txt	./txt/cord-008333-1wepke2o.txt