Yeast expression and DNA immunization of hepatitis B virus S gene with second-loop deletion of α determinant region

Figure 1 Electrophoresis of the fragments of HBV S gene. Lane 1: HB fragment of complete HBV S gene; Lane 2: HB-SD fragment with the second-loop deletion of the α determinant; Lane 3: DNA fragment of HBV S gene from codon 1 to codon 138; Lane 4: DNA fragment of HBV S gene from codon 146 to codon 226; Lane 5: HBY fragments; Lane 6: HB-SDY fragments; Lane M1: DNA marker.

Figure 2 RFLP analysis of recombinant vectors of pHB-SD and pHB-SDY. Lane M1: DNA marker; Lane 1: HB-SD fragment; Lane 2: pHB-SD candidate digested by restriction endonu-clease Hind III and Eco RI; Lane 3: pHB-SD candidate digested by restriction endonuclease Bam HI; Lane 4: pcDNA3 digested by restriction endonuclease Bam HI; Lane 5: pHB-SDY candi-date digested by restriction endonuclease Not I and Eco RI; Lane M2: DNA marker.

Figure 3 Screening integrants and SDS-PAGE analysis. A: Screening integrants using PCR with AOX1 primers. Lane M2: DNA marker; Lane 1: PCR products of Mut⁺ transformant without gene of interest; Lane 2: PCR products of Mut^s transformants with pHBY; Lane 3: PCR products of Mut^s transformants with pHB-SDY; Lane 4: PCR products of pPIC9 alone. Lane M1 were DNA markers. B: Coomassie-stained SDS-PAGE analysis of the recombinant proteins of pHB-SDY and pHBY. Lane M3: protein marker; Lane 5: cell lysate super-natants of yeast alone; Lane 6: yeast transformed with pPIC9; Lane 7: yeast transformed with pHBY; Lane 8: yeast trans-formed with pHB-SDY.

Figure 4 Antigenicity analysis of recombinant proteins using ELISA and immune dot blotting assay. (A) ELISA, (B) immune dot blotting assay.