Schistosoma japonicum: screening of cercariae cDNA library by specific single-chain antibody against SIEA26-28 ku and immunization experiment of the recombinant plasmids containing the selected genes.
To obtain the gene encoding SIEA26-28 ku, which has been proven to be a potential anti-schistosomiasis vaccine candidate, screening Schistosoma japonicum (Sj) cercariae cDNA library with soluble specific single-chain antibody (SIEA26-28 ku-scFv) was performed. A large amount of specific single-chain antibody was harvested through construction of recombinant expression vector pET32a/scFv. The protein was purified and characterized. By using this protein (PET32a-scFv) as a probe, S. japonicum cercariae cDNA library was screened. Two strong positive clones were selected, and their eukaryotic recombinant plasmids were constructed. These genes were named as S. japonicum ribosomal protein S4 (SjRPS4) and S. japonicum ribosomal protein L7 (SjRPL7), respectively. Experiments of mice showed that both SjRPS4 and SjRPL7 DNA vaccines could induce significant immunoprotection. Result of these experiments further proved that the specific single-chain antibody is a very valuable tool in screening of cDNA library to get the corresponding molecules.
Gao
Dong-mei
DM
Department of Parasitology, Xiangya Medical School, Central South University, 110 Xiangya Road, Changsha, 410078, People's Republic of China.
Wang
Shi-ping
SP
He
Zhuo
Z
Fung
Ming-chiu
MC
Liu
Ming-she
MS
Yu
Lu-xin
LX
Chen
Xiu-chun
XC
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D005260:Female; D015723:Gene Library; D007114:Immunization / Q000379:methods*; D007074:Immunoglobulin G / Q000097:blood; D007422:Intestines / Q000469:parasitology; D008099:Liver / Q000469:parasitology; D051379:Mice; D010270:Parasite Egg Count; D010957:Plasmids; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology*; D012269:Ribosomal Proteins / Q000235:genetics / Q000276:immunology; D012549:Schistosoma japonicum / Q000235:genetics / Q000276:immunology*; D012554:Schistosomiasis japonica / Q000276:immunology / Q000469:parasitology / Q000517:prevention & control*; D057127:Single-Chain Antibodies; D014599:Uterus / Q000469:parasitology; D019444:Vaccines, DNA / Q000276:immunology*
D000913:Antibodies, Protozoan; D007074:Immunoglobulin G; D015800:Protozoan Proteins; D012269:Ribosomal Proteins; D057127:Single-Chain Antibodies; D019444:Vaccines, DNA; C031798:ribosomal protein S4; C016700:ribosomal protein L7-L12
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