High-level expression and purification of immunogenic recombinant SAG1 (P30) of Toxoplasma gondii in Escherichia coli.
Surface antigen 1 (SAG1) of Toxoplasma gondii is a good candidate for diagnosis and vaccine development, but recombinant SAG1 produced in Escheichia coli often loses its specific immunogenicity due to the incorrect folding. In the present study, a truncated SAG1 was highly expressed in E. coli as a fusion protein, about 30% of the total protein of the cell lysate. After a simple purification and refolding procedure, purified rSAG1 can be recognized by human Toxoplasma-infective serum, and ELISA kits constructed by rSAG1 can sensitively and specifically detect toxoplasma infection.
Chen
X G
XG
Department of Parasitology, First Military Medical University, Guang ZhoU, 510515, People's Republic of China.
Gong
Y
Y
Hua-Li
Lun
Z R
ZR
Fung
M C
MC
D000818:Animals; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D003001:Cloning, Molecular / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D017510:Protein Folding; D020673:Protein Renaturation; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology*; D014122:Toxoplasma / Q000235:genetics / Q000276:immunology*; D014123:Toxoplasmosis / Q000175:diagnosis / Q000276:immunology; D014170:Transformation, Genetic
D000953:Antigens, Protozoan; D007106:Immune Sera; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C047168:SAG1 antigen, Toxoplasma