Journal: The Journal of Infectious Diseases

Article Title: Genetic Changes at the Glycoprotein Editing Site Associated With Serial Passage of Sudan Virus

doi: 10.1093/infdis/jiv216

Figure Lengend Snippet: Editing site genotype of Sudan virus (SUDV) after serial passage in Vero E6 cells. SUDV was serially passaged in Vero E6 cells at a low multiplicity of infection (MOI) of 0.001 plaque-forming units (PFU)/cell. Ultra-deep sequencing was used to investigate the relative abundance of uridine (U) at the editing site after each passage. A, SUDV was passaged 18 times. B, The first 5 passages were repeated at the same MOI. C, The first 5 passages were repeated at different MOIs (0.01, 0.1, and 1.0 PFU/cell).

Article Snippet: Vero E6 cells (VERO V2008; catalog number NR-596; BEI resources) were grown in minimum essential medium (Life Technologies) containing 2 mM l -glutamine and 1 mM sodium pyruvate (henceforth, referred to as “normal growth medium”) with 10% heat-inactivated fetal calf serum (FCS; Life Technologies) at 37°C with 5% CO 2 .

Techniques: Infection, Sequencing

Journal: The Journal of Infectious Diseases

Article Title: Genetic Changes at the Glycoprotein Editing Site Associated With Serial Passage of Sudan Virus

doi: 10.1093/infdis/jiv216

Figure Lengend Snippet: Editing site genotype and phenotype of Sudan virus (SUDV) after infection in Macaca fascicularis. Eight M. fascicularis were challenged intramuscularly with 100 plaque-forming units of one of 2 SUDV stocks. Four animals received virus that had only been passaged in Vero E6 cell culture 3 times and had a 7-uridine (U) genotype (P3 virus). The other 4 received virus that had been passaged in Vero E6 cell culture 13 times and had an 8-U genotype (P13 virus). A, Nucleic acid was isolated from serum specimens, and ultra-deep sequencing was used to determine the sequence of the editing site in the viral RNA. B, Survival proportions of animals challenged with P3 vs P13 virus. No statistical difference was observed using the log-rank Mantel–Cox test. Abbreviation: NHPs, nonhuman primates.

Article Snippet: Vero E6 cells (VERO V2008; catalog number NR-596; BEI resources) were grown in minimum essential medium (Life Technologies) containing 2 mM l -glutamine and 1 mM sodium pyruvate (henceforth, referred to as “normal growth medium”) with 10% heat-inactivated fetal calf serum (FCS; Life Technologies) at 37°C with 5% CO 2 .

Techniques: Infection, Cell Culture, Isolation, Sequencing