Cloning and expressing of vacA gene related segment with vacuolating activity of Helicobacter pylori
- VernacularTitle:幽门螺杆菌细胞空泡毒素基因毒性片段的克隆与表达
- Author:
Miao LIU
- Publication Type:Journal Article
- Keywords:
Helicobacter pylori;
Vacuolating cytotoxin;
Gene clone;
Expression
- From:
Journal of Chongqing Medical University
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To obtain the antigen of recombinant vacuolating segment of Helicobacter pylori vacuolating cytotoxing by gene-engineering.Methods:The objective gene was cloned into PQE-30,sequenced and expressed using PCR and recombinant techniques.After expression,the antigenicity of recombinant protein was analyzed by Western blotting.Results:DNA sequence analysis identified that the sequence was integrated.Compared with standard strain AF361700,1.5% of the cloned gene was mutated,one amino acid radicle was changed.SDS-PAGE revealed the molecular weight of objective recombinant protein was 27 000 as it was predicted;the level of expression product was over 30%.Western blotting results showed the objective protein could be recognized by anti-serum against HP.Conclusion:This recombinant protein has perfect antigenicity,so it may be used as a protective antigen in preventing HP infection.
