Objective: To clone and identify the Mycobacterium bovis BCG gene for signal peptide of antigen 85-B.
Methods: Using a polymerase chain reaction technique, the signal peptide sequence (position 94 to 211) of antigen 85-B was technique, which is one of the major protein secreted by Mycobacterium bovis BCG (Bacille-Calmette-Guerin). The gene of 117bp fragment was cloned into the SacI and EcoRI sites of pBluecript SK, and its sequence was determined by using T7DNA polymerase kit.
Results: The sequence analysis showed that the sequence of 117bp signal peptide was not misincorporated and in 3' end understream of signal peptide the 12 multi cloning sites for foreign gene could be provided by the recombinant plasmid.
Conclusions: The signal sequence of BCG antigen 85-B was cloned into the pBluescript plasmid. The recombinant plasmid was called BCG-S01. According to the genetic information a secreted protein from M. bovis BCG could be utilized in making a useful vaccine vehicle to produce and secrete a vaccinal protein from M. bovis BCG living cells.