Expression of the recombinant fusion protein CP15-23 of Cryptosporidium parvum and its protective test.
The CP15 and CP23 surface proteins on the sporozoite of Cryptosporidium parvum are major protective antigens. The recombinant plasmid pET28-15-23 was constructed based on the plasmids pMD18-T-15 and pMD18-T-23 with two pairs of specific primers using DNA recombinant technique. In the primers, a synthetic linker sequence encoding a peptide (G-G-S) was designed. After identification, the recombinant plasmids were transformed to component cells of Escherichia coli BL21 (DE3). The positive strain containing the recombinant plasmid could express a specific fusion protein (CP15-23, MW approximately 25 kDa) induced by IPTG. The fusion protein could be recognized by the positive serum of mice infected with Cryptosporidium parvum oocysts specifically. The BALB/c mice were immunized with 80 microg of CP15-23 protein 4 times at 2 week intervals. The mice produced specific antibodies that responded to the lysate of Cryptosporidium parvum oocysts and could prevent Cryptosporidium parvum infection. The results indicated that the recombinant fusion protein CP15-23 would be used as a candidate antigen to prevent cryptosporidiosis.
Hong-Xuan
He
H
State Key Lab of Biomembrane and Membrane Technology, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, P R. China.
Lei
Chen
C
Cheng-Min
Wang
W
Kai
Zhou
Z
Yi
Tang
T
Xi-Ming
Qin
Q
Ming-Xing
Duan
D
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000918:Antibody Specificity; D001483:Base Sequence; D003457:Cryptosporidiosis / Q000276:immunology / Q000517:prevention & control*; D016785:Cryptosporidium parvum / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D016054:DNA, Protozoan / Q000235:genetics; D015870:Gene Expression; D017125:Genes, Protozoan; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D036103:Nanotechnology; D010957:Plasmids / Q000235:genetics; D015800:Protozoan Proteins / Q000235:genetics* / Q000276:immunology; D016052:Protozoan Vaccines / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology
D000913:Antibodies, Protozoan; C094890:CP15 protein, Cryptosporidium parvum; D016054:DNA, Protozoan; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; D011993:Recombinant Fusion Proteins