Comparative Evaluation of T-Cell Immune Response to BTV Infection in Sheep Vaccinated with Pentavalent BTV Vaccine When Compared to Un-Vaccinated Animals

Table 1

Oligonucleotide primer sequences and optimal amplification conditions of RT-qPCR reaction for cytokines, β-actin and BTV.

Set	Primer name	Primer sequence (5′-3′)^a	Product size (base)	Primer length (base)	Anneling T° (°C)	Primer binding position	Accession number
1	IFN-α FP	ACCTTCCAGCTCTTCAGCACAGA	187	23	60.0	190–212	AY802984
	IFN-α RP	TGTGGAAGTGTTTCCTCACAGCCA		24	60.4	353–376
2	IFN-γ FP	CTTGAACGGCAGCTCTGAGAAACT	91	24	59.0	367–390	X52640
	IFN-γ RP	ATTGATGGCTTTGCGCTGGATCTG		24	60.0	434–457
3	IL-2 FP	CAAACTTCTAGAGGAAGTGCTAGAT	76	25	59.7	192–216	X60148
	IL-2 RP	GTCCATTGAATCCTTGATCTCTCT		24	57.2	244-267
4	IL-6 FP	ACTGCTGGTCTTCTGGAGTA	100	20	57.3	364–383	X68723
	IL-6 RP	TTCTGATACTGCTCTGCAACTC		22	58.4	442–463
5	IL-12 FP	TCTTCACAGACCAAACCTCAGCCA	111	24	60.0	863–886	NM001009438
	IL-12 RP	ACACAGATGCCCATTCACTCCAGA		24	60.0	950–973
6	TNF α FP	TGGGCCAACTCCCTCTGTTTATGT	163	24	59.5	2195–2218	EF446377
	TNF α RP	AGTTTGTGTCTCCCAGGACACCTT		24	59.9	2291–2314
7	β-actin FP	TGAAGATCCTCACGGAACGTGGTT	84	24	59.9	305–328	AF129289
	β-actin RP	AGCAGAGCTTCTCCTTGATGTCAC		24	58.5	365–388
8	Seg5 (NS1) FP	GCAGCATTTTGAGAGAGCGA	101	20	59	169–188	JQ740775
	Seg5 (NS1) RP	CCCGATCATACATTGCTTCCT		21	58	249–269

^aPrimers were designed based on sequences of ovine cytokine genes from GenBank database.