FIG. 1.

MPT83 antigen expression following MPT83 DNA or MPT83 RNA transfection. Western blot analysis of MPT83 expression by J774 (a)- or BHK (b)-transfected cells. Whole-cell extracts were prepared from two independent J774 cell lines stably transfected with CMV4.83 or BHK cells transiently transfected with MPT83 RNA prepared according to the protocol described in Materials and Methods. Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, blotted onto nitrocellulose membranes, and incubated with the specific anti-MPT83 monoclonal antibody 12/6/1. Also shown are kinetics of MPT83 expression, in vivo, following intramuscular injection with MPT83 RNA (c) or with MPT83 DNA (d). Total RNA was purified from the injected muscles and reverse transcribed, and the cDNA was amplified by PCR using MPT83-specific primers. The plasmids used were pCMV4.83 (lanes 1), pCMV3.83 (lanes 2), and pMGD20.83 (lanes 3) (24). The MPT83 RNA was synthesized in vitro using the plasmid pSinRep5.83 as the DNA template and the SP6 transcription kit as described in Materials and Methods.