| Summary: | 碩士 === 大仁科技大學 === 生物科技研究所 === 96 === Helicobacter pylori ( HP ) is a Gram-negative bacterium. According to previous studies, the human chronicgestritis, gastric ulcer, duodenal ulcer , gastric cancer and other stomach cancer is strongly related to infection of HP. Immunoglobulin Y ( IgY ) is the major antibody from chickens. The specific IgY antibody comes after immunization with these recombinant proteins derived from the virulent factor of HP. The advantages of IgY are high stability, easy to be produced and mass production as well. The objective of this study is to amplify gene fragment of CagA which is a virulence factor of HP by polymerase chain reaction ( PCR ) techniques. For cloning cagA gene, pGEX6p-1 was used as a vector to construct the recombinant plasmid. The recombinant plasmid was transformed into E. coli DH5α for protein expression. By using affinity column the, expression of fusion protein ( 165 kDa ) could be purified. By using Western blotting assay, we took the rabbit anti HP polyclonal antibody and IgY against the CagA recombinant protein from the immunized chickens for analyzing the antigenicity and immunogenic. The resulting data revealed that both kinds of specific antibodies described above could strongly reacted to the CagA recombinant protein ( 165 kDa ) in terms of antigenicity and immunogenicity. However, this study proved that the lgY had titers as high as 1024X. In vitro, the bacterial growth inhibition test was found that 15 mg/ml of IgY could completely inhibit the growth of 4.5×104 CFU/mL of H. pylori. Since approximately 91.39 mg of IgY from each yolk can be obtained, it could be feasible for mass production of this IgY antibody. It seems to be true that a new approach for the treatment of gastric ulcer could be developed in the future.
|
|---|
