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. 2007 Apr 18;1(2):147–157. doi: 10.1016/j.chom.2007.03.003

Figure 2.

Figure 2

Rescue of rsT3D and rsT3D/T1LS1

(A) Electropherotypes of T1L, T3D, rsT3D, and rsT3D/T1LS1. Viral dsRNA was extracted from purified virions and electrophoresed in an SDS-polyacrylamide gel, followed by ethidium bromide staining to visualize viral gene segments. Size classes of gene segments (L, M, S) are indicated.

(B) Recombinant viruses contain a novel mutation in the L1 gene. The single nucleotide difference in L1 unique to rsT3D and rsT3D/T1LS1 is shown in the alignment as an asterisk. The G→A substitution at position 2205 is a signature change engineered into the cloned T3D L1 cDNA used for marker rescue.

(C) Sequence analysis of L1 gene RT-PCR products from rescued reoviruses. A fragment of the L1 gene was amplified by one-step RT-PCR performed using viral dsRNA extracted from purified virions of T3D, rsT3D, and rsT3D/T1LS1. Products were subjected to direct sequence analysis and compared to the L1 sequence of T3D. Shown are sequence chromatograms demonstrating G→A substitution at position 2205 of the rsT3D and rsT3D/T1LS1 L1 genes.