Journal: PLoS Biology
Article Title: The dinucleotide composition of the Zika virus genome is shaped by conflicting evolutionary pressures in mammalian hosts and mosquito vectors
Figure Lengend Snippet: Fitness comparison between WT and mutant ZIKV in human and mosquito cells. Human A549 cells, A549 ZAP knockout cells, and AP-61 mosquito cells were infected with equal RNA concentrations of WT ZIKV mixed with either the SCR (A) , CpG-high viruses (CpG_1.0 and CpG_max) (B, C), or UpA_max (D) . Virus was passaged to new cells twice, before total RNA was isolated. ZIKV RNA was amplified with RT-PCR and digested with BsaI (A), SalI (B, C), or HeaIII (D) restriction enzymes to distinguish between WT and the respective mutant viruses. DNA gel images display the largest most distinctive products from each digestion. For more information, see S3 Fig . From left to right images display the digested DNA of the indicated individual viruses, followed by 2 independent experimental repeats performed with separately rescued virus populations (numbered 1 and 2). RT-PCR, reverse transcription PCR; SCR, scrambled control virus; WT, wild-type; ZAP, zinc-finger antiviral protein; ZIKV, Zika virus.
Article Snippet: The resulting DNA amplicons from the comparisons between WT and SRC were digested with BsaI-HFv2 (New England Biolabs), WT and CpG_1.0 or CpG_max with SalI-HF (New England Biolabs) and WT and UpA_max with HaeIII (Invitrogen) and ran on 2% agarose gels.
Techniques: Mutagenesis, Knock-Out, Infection, Isolation, Amplification, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction