Schematic
Figure 1 (a): Schematic steps followed to obtain the fused proteins based on gene fragment MPT-64 CE15 peptide (42 bp located at the position 174-187 aa of the MPT-64) or MPT-6491L-205A sequence (348bp) insertion between MT10.3(1M-40S) (120 pb) and MT10.3 (41S-96) (171bp), respectively. Fragments were amplified by PCR, from M. tuberculosis DNA, with homologous primers containing enzymes restrictions sites. As the CE15 fragment was not able to be ligated at pQE80L expression vector, the successful construction containing pQE80L-MT10.3 (1M-40S):MPT-64(91L-205A):MT10.3(41S-96) (F2) was used. The fragment MPT-64(91L-205A) was digested, and into SacI/SalI predigested F2 was inserted the same enzymes pre-digest CE15 PCR fragment generating the pQE80L-MT10.3 (1M-40S):CE15 (173G-187A):MT10.3 (41S-96) (F1).