Article Figures & Data
Figures
- Fig. 1.
A, Southern blot hybridization of PCR products obtained from the 1955 poliovaccine with the primer set 2902/2573. Lane 1, 10 μl of poliovaccine; Lane 2, 2 μl of poliovaccine; Lanes 3 and 5, empty; Lane 4, mock DNA extraction; Lane 6, SV40 DNA positive control. B, Southern blot hybridization of PCR products obtained from 1996 poliovaccines with the primer set 2902/2573. Lanes 1–6, 10 μl of oral poliovaccine; Lanes 7–12, 10 μl of parenteral poliovaccine; Lane 13, mock DNA extraction; Lane 14, water; Lane 15, SV40 DNA positive control. C, scheme showing the PCR strategy used to sequence the SV40 genome from the poliovaccine. The inner circle represents the first set of PCRs amplifying fragments of an average size of 330 bp. Only 14 of the 17 amplified fragments are shown due to space limitations. The empty space between fragments represents the region where the primers anneal. The middle and outer circles represent different combinations of the above cited primers to read the SV40 sequence in the region where the primers anneal and to confirm the polymorphisms detected.
- Fig. 2.
A, products of PCR amplification with the R1/R2 set of primers from two different lots of 1955 poliovaccines (ethidium bromide staining of 2% agarose gel). Each lot was tested in triplicate: −, H2O negative control, +, SV40 DNA positive control. B, the alignment of the sequence of SV40-776 origin of replication and early region promoter with the sequence obtained from the poliovaccine 028846B shows a deletion of one of the two 72-bp enhancers in the poliovaccine. •, homology; −, deletion.
- Fig. 3.
Top, infection of TC-7 cells with nonarchetypal (2 72bP) SV40 at a MOI of 10−2 (A) and a MOI of 10−4 (C). Infection of TC-7 cells with archetypal (1 72bP) SV40 at MOIs of 10−2 (D), 10−3 (E), and 10−4 (F). Uninfected TC-7 cells (B). The pictures were taken 14 days after infection. (×400). Bottom, Tag immunostaining of TC-7 cells infected with nonarchetypal (2 72bP) SV40 at MOIs of 10−2 (A) and 10−4 (C). TC-7 cells infected with archetypal (1 72bp) SV40 at MOIs of 10−2 (D), 10−3 (E), and 10−4 (F). The staining was done on day 14 after the infection. Uninfected TC-7 cells (B). (×200).
Tables
- Table 1
Polymorphisms found in PV028846B and their location in the SV40 genome
nt no. SV40-776 Vaccine isolate 36 G A 107–178 del. 72 bp 732 T A 747 C A 1756 A C 1939 T A 2239 T G 2716–2721 TGGGAG del. 6 bp 2751 A G 2757 G A 2766–2771 ATTATG del. 6 bp 2795 — TGAGGGCTG 2813 C T 2851 T A 2908–2913 TCATCA del. 6 bp 2950 A G 3117 T C 3727 C G 3755 A G 3873 C T 4834 G A 4839 C T 4879 C T
Volume 59, Issue 24
