Studies on immunogenicity of Schistosoma thioredoxin Ⅰ. Construction and identification of DNA vaccine of Schistosoma japonicum thioredoxin
- VernacularTitle:血吸虫硫氧还蛋白免疫原性研究 Ⅰ日本血吸虫硫氧还蛋白DNA疫苗的构建和鉴定
- Author:
Jianping CAO
;
Haibo HAN
;
Shuxian LIU
- Publication Type:Journal Article
- Keywords:
Schistosoma japonicum;
Thioredoxin;
Eukaryon expression vector;
DNA vaccine;
Mice;
Immunization
- From:
Chinese Journal of Schistosomiasis Control
1989;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo construct and identify a DNA vaccine of Schistosoma japonicum (Chinese strain) thioredoxin. MethodsAccording to a cDNA sequence of S.japonicum (Philippine strain) thioredoxin, a couple of primers was designed with the BamH I restriction endonuclease site introduced in forward primer with ATG as start condon and EcoR I in reverse primer with TCA as termination codon. The gene encoding S.japonicum (Chinese strain) thioredoxin (SjcTrx) was amplified by RT-PCR using total RNA of schistosome as the template. The PCR products and the pcDNA3 plasmids were digested by restriction endonucleases BamH I and EcoR I. The target DNA fragment were purified and cloned properly into the eukaryotic expression vector of pcDNA3. The recombinant plasmids were then transformed into competent E.coli JM109 and identified by endonucleases digestion, PCR, agarose gel electrophoresis and sequencing. ResultsThe RT-PCR product was around 334 bp judged by agarose gel electrophoresis. The same fragments were obtained by restriction enzyme digestion from the recombinant plasmid and PCR with the plasmid DNA as a template. The recombinant plasmid, designated pcDNA3-SjcTrx, was sequenced and shown to be 97% and 43% identical in deduced amino acid sequences to that of S.japonicum (Philippine strain) and S.mansoni thioredoxin, respectively. ConclusionThe S.japonicum thioredoxin DNA vaccine had been constructed successfully, and further studies will be made in different animal models for its immunogenicity.
