Identification and expression of the DNA vaccines in vitro. (A) Restriction enzyme analysis of the 3 multi-epitope DNA vaccines, with digestion by BamH I and EcoR I,follwed by 1% agarose electrophoresis. Two bands (about 5,000 bp and 500 bp) were observed for all 3 recombinant plasmids. Lane 1, pJW4303-MEG1; lane 2, pJW4303-MEG2; lane 3, pJW4303-MEG3; M, Trans 2K Plus marker. (B) Western blot analysis of DNA vaccine protein expression in vitro. 293T cells were transiently transfected with pJW4303-MEG1, pJW4303-MEG2, or pJW4303-MEG3. Expression of the proteins was identified by Western blot using rabbit anti-MEG1.E polyclonal antibody as primary antibody. A 16 kDa band was observed for both pJW4303-MEG1 and pJW4303-MEG2, but no signal was detected for pJW4303-MEG3 or the control. Lane 1, pJW4303-MEG1; lane 2, pJW4303-MEG2; lane 3, pJW4303-MEG3; lane 4, negative control. Protein marker beside the film used to illustrate protein sizes was cut from the PVDF membrane.