Table 1.

Overlapping segments amplified from pDAFL plasmid containing Daniels strain of Theiler’s murine encephalomyelitis virus (TMEV).

Segment	Location in TMEV genome	pcDNA3.1 His A cloning sites
TMEV 1	1038–1596	EcoRI 5′; XbaI 3′
TMEV 2	1536–2097	BamHI 5′; XbaI 3′
TMEV 3	2033–2596	EcoRI 5′; XbaI 3′
TMEV 4	2534–3099	BamHI 5′; XbaI 3′
TMEV 5	3031–3597	EcoRI 5′; XbaI 3′
TMEV 6	3532–4095	EcoRI 5′; XbaI 3′
TMEV 7	4030–4596	EcoRI 5′; XbaI 3′
TMEV 8	4531–5094	BamHI 5′; XbaI 3′
TMEV 9	5032–5596	EcoRI 5′; XbaI 3′
TMEV 10	5533–6096	EcoRI 5′; XbaI 3′
TMEV 11	6031–6597	EcoRI 5′; XbaI 3′
TMEV 12	6532–7095	EcoRI 5′; XbaI 3′
TMEV 13	7033–7597	EcoRI 5′; XbaI 3′
TMEV 14	7531–7971	EcoRI 5′; XbaI 3′
TMEV L*	1064–1547	EcoRI 5′; XbaI 3′

Fifteen overlapping segments were amplified from the pDAFL plasmid containing the TMEV genome using high-fidelity polymerase chain reaction. Segments were then digested with the specified restriction enzyme as defined by primer sequences and inserted into the pcDNA 3.1 His A vector to accommodate transfection into the Cos-7 cell line. The TMEV sequences overlap by 30 base pairs to ensure that all potential CD8+ T-cell epitopes are processed for antigen presentation.

^*Note that the L segment encodes a protein from an alternative reading frame important to TMEV infection.