id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-017824-0pinevfc	Tekes, Gergely	Vaccinia Virus-Based Reverse Genetics for Feline Coronaviruses	2015-09-10		.txt	text/plain	3190	253	57	The main focus of this chapter is the vaccinia virus-based reverse genetic system for FCoVs. Here we present protocols for (1) the generation of a full-length cDNA clone, (2) the manipulation of the FCoV genome, and (3) the rescue of recombinant FCoVs. The establishment of a reverse genetic system for feline coronaviruses (FCoVs), which allows to modify the entire coronaviral genome, was successfully achieved for the fi rst time in 2008 [ 1 ] . Regardless of the applied strategy, viral RNA serves as a starting material for the generation of all FCoV-sequence containing plasmids (sections "First Step", 3.1.1.3, "Generation of plasmids suitable for vaccinia virus-mediated homologous recombination"). In order to integrate the full-length FCoV cDNA, fragments 1-8 are introduced via four rounds of vaccinia virus-mediated homologous recombination using GPT as a positive and a negative selection marker.	./cache/cord-017824-0pinevfc.txt	./txt/cord-017824-0pinevfc.txt