id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-334947-pa0p5dif	Rozen-Gagnon, Kathryn	Alphavirus Mutator Variants Present Host-Specific Defects and Attenuation in Mammalian and Insect Models	2014-01-16		.txt	text/plain	8887	415	44	Since residue 483 is conserved among alphaviruses, we examined the analogous mutations in Sindbis virus (SINV), which also reduced polymerase fidelity and generated replication defects in mosquito cells. To estimate the population diversity of variants by deep sequencing, cDNA libraries were prepared by Superscript III from RNA extracted from virus generated in BHK-21 or C6/36 cells, and the viral genome was amplified using a high fidelity polymerase (Phusion) to generate 5 overlapping amplicons 2-3 kb in length. To address the possibility that the fitness cost of defective replication, observed in mosquito cell culture, would favor the reversion of these mutant polymerases to wildtype, we deep sequenced virus from the body of an individual mosquito that presented the median titer from each group.	./cache/cord-334947-pa0p5dif.txt	./txt/cord-334947-pa0p5dif.txt