Molecular cloning, expression and vaccination of a novel gene Sj-MA of Schistosoma japonicum.
In order to obtain new gene and to develop the new vaccine candidate of immunoprotection against Schistosoma japonicum (Sj), Sj adult worm cDNA library was screened with anti-sera to soluble male adult worm antigen and resulted in the discovery of a novel gene designated as Sj-MA. Sequence analysis showed that Sj-MA as a complete cDNA contains one open reading frame. It was deduced to contain 249 amino acid residues and encode a 28.8 kD soluble protein with plenty of phosphorylation sites, supposing its action in signal transduction. Furthermore, Sj-MA cDNA was cloned into a prokaryotic expression vector pGEX-5X to construct the recombinant plasmid which was transformed and highly expressed in E. coli as a 54.8 kD glutathione-S-transferase (GST) fusion protein. The fusion protein rSj-MA/GST could be recognized with both anti-male adult worm sera and anti-GST sera in Western blotting. Mice vaccinated with the fusion protein revealed significant worm reduction rate of 34.29% (P<0.001), compared with the control groups. Taken together, the novel gene Sj-MA can be expressed in E. coli as a fusion protein that can elicit immunity against Schistosoma japonicum, suggesting its potential as a new vaccine candidate.
Chen
Yu-Xiao
YX
Department of Immunology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
Yi
Xin-Yuan
XY
Zeng
Xian-Fang
XF
Yuan
Shi-Shan
SS
Zhang
Shun-Ke
SK
McReynolds
Larry
L
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D015801:Helminth Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D007074:Immunoglobulin G / Q000097:blood; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012549:Schistosoma japonicum / Q000235:genetics* / Q000276:immunology; D012555:Schistosomiasis mansoni / Q000276:immunology / Q000517:prevention & control; D017422:Sequence Analysis, DNA; D014611:Vaccination
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018076:DNA, Complementary; D015801:Helminth Proteins; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; C482321:Sj-MA protein, Schistosoma japonica; D005982:Glutathione Transferase