Construction of hpaA gene from a clinical isolate of Helicobacter pylori and identification of fusion protein

Figure 1 The target fragment of hpaA gene amplified from H. pylori strain Y06 (1: 100 bp DNA marker; 2: blank control; 3 and 4: the target fragment amplified from H. pylori strain Y06 DNA).

Figure 2 Homology comparison of H. pylorihpaA gene nucleotide sequences ([1]-[6]: the reported sequence from GenBank (No. X92502, strain 11637; No. NC000915, strain 26695; No. X61574, strain 8826; No. NC_000921, strain J99; No. AF479028, strain CH-CTX1; No. U35455, strain CCUG 17874); [7]: the sequencing result of H. pylori strain Y06 hpaA gene. outline region: oligonucleotide primer sequences; square frame: start code and end code).

Figure 3 Homology comparison of the putative amino acid sequences of hpaA gene ([1]-[6]: the reported sequence from GenBank (No. X92502, strain 11637; No. NC000915, strain 26695; No. X61574, strain 8826; No. NC_000921, strain J99; No. AF479028, strain CH-CTX1; No. U35455, strain CCUG 17874); [7]: the sequencing result of Hp strain Y06 hpaA gene).

Figure 4 Expression of HpaA gene in pET32a-hpaA-BL21(DE3) induced by IPTG with different dosages shown in SDS-PAGE. (1: protein marker; 2: non-induced; 3-5: induced with 0.1, 0.5, 1 mmol•l^-1 of IPTG, respectively; 6: the bacterial precipitate induced with 1 mmol•l^-1 of IPTG; 7: the bacterial supernatant induced with 1 mmol•l^-1 of IPTG).

Figure 5 Western blot result of commercial rabbit antibody against whole cell of H. pylori and HpaA fusion protein. (1 and 2: 20 μL and 40 μL of expressed HpaA product induced with 0.1 mmol•l^-1 of IPTG, respectively).