A POOL OF RECOMBINANT HELICOBACTER PYLORI VACUOLATING CYTOTOXIN FRAGMENTS FROM E.COLI FOR VACCINE

摘要

A method for preparing a vaccine comprising recombinant Helicobacter pylori vacuolating cytotoxin(VacA) fragments produced from Escherichia coli is provided to preserve an epitope of an VacA antigen produced from Escherichia coli, thereby obtaining a neutralizing antibody against a natural VacA antigen through animal immunization. A method for preparing a vaccine comprising recombinant Helicobacter pylori vacuolating cytotoxin(VacA) fragments produced from Escherichia coli comprises the steps of: designing a primer by using pET41b, VacA gene of ATCF49503 and VacA gene of ATCC 43504; amplifying the VacA genes through PCR(polymerase chain reaction); digesting the pET 41b and PCR product with recombinant enzymes XhoI and NdeI; ligating the digested products at 20 deg. C for 30 minutes and transforming the digested products to a cell ECOS 101 to produce a recombinant Helicobacter pylori vector pVAC 953; transforming Escherichia coli with the pVAC 953; culturing the transformed E. coli in a medium containing IPTG(isopropyl-dithiogalactopyranoside) at 20 deg. C to induce protein expression; recovering the protein-expressed E. coli and reacting with an enzyme such as lysozyme; pulverizing the cells, obtaining the supernatant and subjecting the supernatant to column chromatography; filtering and concentrating the obtained recombinant VacA protein; and mixing the concentrated antigen with an adjuvant such as CpG-ODN having unmethylated cytosine and guanine motifs.