Immunogenicity and Protective Efficacy against Murine Tuberculosis of a Prime-Boost Regimen with BCG and a DNA Vaccine Expressing ESAT-6 and Ag85A Fusion Protein

Table 1

Primers and thermal cycle parameters for cloning of M. tuberculosis antigens.

Protein Names	Primer names and sequences (5′-3′)	Cycle parameters
ESAT-6	ESAT6F: GGATCCATGACAGAGCAGCAGTG; ESAT6R: GCTGCCGCCACCGCCGCTTCCGCCACCGCCGCTTCCACCGCCACCTGCGAACATCCCAGTGACGTTGCCTTC	95°C 5 min; 94°C 45 s, 60°C 45 s, 72°C 50 s, 30 cycles; 72°C 10 min
Ag85A	Ag85AF: GGTGGCGGTGGAAGCGGCGGTGGCGGAAGCGGCGGTGGCGGCAGCGCATTTTCCCGGCCGGGCTTG; Ag85AR: GGAATTCTGTTCGGAGCTAGGCGCCCTGGG	95°C 5 min; 94°C 45 s, 60°C 45 s, 72°C 50 s, 30 cycles; 72°C 10 min
r685A	ESAT6F; Ag85AR	95°C 5 min; 94°C 1 min, 60°C 1 min, 72°C 90 s, 30 cycles; 72°C 10 min