Gene cloning and characterization of hopX outer membrane protein from Helicobacter pylori
- VernacularTitle:幽门螺杆菌外膜蛋白hopX基因克隆表达及生物信息学分析
- Author:
Xiaoli JU
;
Shihe SHAO
;
Weibin HAO
- Publication Type:Journal Article
- Keywords:
Helicobacter pylori;
Sequencing;
Biological information
- From:
Chinese Journal of Immunology
1985;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To clone the outer membrane protein hopX gene of Helicobacter pylori(Hp)and to perform sequencing and analysis of biological information.Methods:Polymerase chain reaction(PCR)was used to amplify the hopX gene from Hp chromosomal DNA.Then the target gene was digested by restricted endonuclease enzyme of BamH I,and inserted into the prokaryotic expression vector pQE30 digested by corresponding restricted endonuclease enzyme.The recombinant vector was used to select and transform for nucleotide sequence analysis.The biological property at the amino acid level was analyzed by Omiga 2.0 and Antheprot v 5.0.The transformant colony was induced with IPTG and the fusion protein was analyzed by SDS-PAGE and Western blot.Results:The recombinant plasmid was constructed.DNA sequence analysis showed the sequence of hopX was 1 284 bp.The homology of the strains in nucleotide acid was 96%～97%.Their homogeneity in the amino acids was 97%～99%.We get a GeneBank accession number EF208122.Omiga 2.0 software predicted its relative molecular mass(Mr.)was 47 kD and possessed good antigencity.The expressed product contained about 37% of total somatic proteins and Western blot method showed good antigenicity of the recombinant protein.Conclusion:A confirmed gene hopX has been obtained,providing a good foundation for recombination,expression and related study.The corresponding peptide of the gene performed the structural characteristics of some typical antigen molecules,which suggest that it might be a novel vaccine candidate.
