Nucleotide and amino acid changes in uncloned and cloned ChimeriVax-DEN viruses

VirusVersionMutations detected at sequenced passagesa
ChimeriVax-DEN22000UnclonedP2, noneb; P5, noneb; P10, A1716T (KE-246M), G3287A (VNSI-275M)b
Clone AP2, G7734T (RNS5-29M)c; P6, G7734T (RNS5-29M)b; P8, A515/C (MprM-12/L), C2135/G (QE-386/E), G7734T (RNS5-29M)b
Clone BP2, A6643T (silent)c; P6, A6643T (silent)b; P9, G517/C (MprM-12/I), A6643T (silent)c; P14, G517C (MprM-12I), A6643T (silent), C7427T (LNS4B-177F)c; P18, G517C (MprM-12I), A6643T (silent), C7427T (LNS4B-177F)b
ChimeriVax-DEN22001UnclonedP2, noneb; P15, A515C (MprM-12L), G1321A (silent), A1590G (KE-204R), A4046T (INS2A-176L), T5485C (silent), C6888T (A2K-20V), C7427T (LNS4B-177F), G9094/T (silent), C10307T (silent)b
Clone AP6, noneb; P7, noneb; P10, T551G (LprM-24V), G1730T (VE-251F)b; P20, T551G (LprM-24V), G1730T (VE-251F)b
Clone BP3, nonec; P6, A1030G (silent), C1789T (silent)d; P15, A1030G (silent), A1590G (KE-204R), C1789T (silent), T2444C (silent), C7443T (ANS4B-182V), A8677G (silent)b
ChimeriVax-DEN3UnclonedP2, noneb; P5, nonec; P10, C564A (AprM-28D), T2561C (YNS1-35H)c; P15, C564A (AprM-28D), T2561C (YNS1-35H), A6502/G (silent), A7231/G (INS4B-113/M)b
Clone AP3, C6607T (silent)c; P6, C6607T (silent)b; P7, C6607T (silent)b; P10, C6607T (silent)c; P15, C6607T (silent), C7859/T (silent)c,e; P20, C6607T (silent), C7421T (LNS4B-177F), C7859/T (silent)b
ChimeriVax-DEN4UnclonedP2, noneb; P15, noneb
Clone BP6, noneb; P7, noneb; P10, noneb; P20, noneb
ChimeriVax-DEN1UnclonedP2, noneb; P5, A1590/G (KE-204/R), G1730/T (VE-251/F)c; P15, A1590G (KE-204R), G1730/T (VE-251/F), A7237/G (INS4B-113/M), C7466/T (PNS4B-190/S)b
Clone AP3, G1730T (VE-251F), C2282A (LE-435I)c; P7, G1730T (VE-251F), C2282A (LE-435I)b
Clone B, DP3, G1730T (VE-251F)c; P7 or P6, G1730T (VE-251F)b
Clone CP3, G1912T (EE-311D), G2030T (VE-351L)c; P6, G1912T (EE-311D), G2030T (VE-351L)b
Clone EP3, A1590G (KE-204R), A3952T (silent)c; P6, A1590G (KE-204R), A3952T (silent)b
Clone FP3, C788T (silent), A1590G (KE-204R)d
Clone GP3, G1730T (VE-251F)d
Clone HP3, G1912T (EE-311D), G2030T (VE-351L)d
Clone IP3, A1590G (KE-204R)d
Clone JP3, noneb; P6, noneb; P7, noneb; P10, A1590G (KE-204R)b; P20, A1590G (KE-204R), G6966/T (SNS4B-23/I), G7190/A (VNS4B-98/I)b
  • a The passage number is shown first; any mutation(s) detected follow the passage number. Nucleotide changes at indicated positions (numbering from the beginning of the genome) and amino acid changes (in parentheses) at indicated protein residues [from the N terminus of each protein; amino acid numbering of YF proteins as in references 3 and 17] are shown. A heterogeneous signal is indicated by a slash. Data for clones after the first and/or last plaque purifications are in bold type.

  • b Full genome sequenced.

  • c Sites of mutations found after last plaque purification (in samples after first plaque purification) or at a later genetic stability passage were sequenced only. It can be assumed that the full genome (or fragment I) sequence is known.

  • d Only fragment I sequenced, including the entire structural protein coding region.

  • e In independent passages of the DEN3 clone A virus to P15 from a Master Seed (large-scale manufacturing P8), accumulation of A1584G (KE-202R; identical to the E-204 mutation in the DEN1 and DEN2 chimeras) and C7437T (ANS4B-182V) mutations was documented, in addition to the silent nt 6607 change (F. Mitchell, data not shown).