Cloning, expression and protective efficacy evaluation of radiation sensitive protein 23 (RAD23) from Schistosoma japonicum.
- Author:
Changjian LI
;
Min ZHANG
;
Yang HONG
;
Yanhui HAN
;
Xiaodan CAO
;
Hongxiao HAN
;
Zhiqiang FU
;
Chuangang ZHU
;
Ke LU
;
Hao LI
;
Jiaojiao LIN
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Helminth;
blood;
Blotting, Western;
Cloning, Molecular;
DNA Repair Enzymes;
genetics;
metabolism;
DNA, Complementary;
Enzyme-Linked Immunosorbent Assay;
Escherichia coli;
Genetic Vectors;
Helminth Proteins;
genetics;
immunology;
Immunoglobulin G;
blood;
Mice;
Mice, Inbred BALB C;
Recombinant Proteins;
genetics;
immunology;
Schistosoma japonicum;
genetics;
metabolism;
Schistosomiasis japonica;
prevention & control;
Vaccines;
immunology
- From:
Chinese Journal of Biotechnology
2014;30(11):1669-1678
- CountryChina
- Language:Chinese
-
Abstract:
Radiation sensitive protein 23 (RAD23) is a nucleotide excision repair (NER) protein that plays an important role in Ubiquitin-proteasome pathway (UPP). Schistosoma japonicum radiation sensitive protein23 (SjRAD23) cDNA sequences were amplified by PCR and cloned into pET28a (+) vector to construct recombinant expression plasmid pET28a(+)-SjRAD23. The recombinant protein was expressed as both inclusion bodies and the supernatant in Escherichia coli BL21 (DE3) cell. Immunofluorescence observation shows that SjRAD23 was mainly distributed on the tegument surface of the worms. ELISA assay reveals that specific IgG, IgG1 and IgG2a antibodies could be detected in the sera of rSjRAD23 immunized mice. Western blotting analysis shows that the recombinant SjRAD23 could be recognized by serum specific to soluble adult worm antigen of S. japonicum. BALB/c mice vaccinated with rSjRAD23 combined with 206 adjuvant revealed 35.94% worm reduction and 40.59% liver egg reduction when compared with that of the adjuvant control
