Gene cloning, expression and vaccine testing of Schistosoma japonicum SjFABP.
A 600 bp DNA fragment was amplified by PCR from an adult Schistosoma japonicum cDNA library. Sequence analysis confirmed that this fragment contained an S. japonicum Chinese mainland strain fatty acid binding protein (Sj14FABP) gene. This gene was subsequently expressed in Escherichia coli (E. coli) and in Baculovirus/silkworm systems. The recombinant protein from E. coli was a 41 kDa GST fusion protein (rSj14/GST), which could be purified by glutathione agarose affinity chromatography, with a yield of 25 mg/L E. coli culture. The recombinant protein from the Baculovirus/silkworm system was an 18 kDa fusion protein (rSj14/His), which could be purified by Ni-NTA resin chromatography column with a yield of 3.5 mg per silkworm larva. Both rSj14/GST and rSj14/His could be recognized by S. japonicum-infected mouse sera and anti-rSj14/GST mouse sera in Western blotting. The purified recombinant protein was immunogenic in mice, rats and sheep, and 34.3%, 31.9% and 59.2% worm reductions, respectively, were obtained in vaccinated Kunming mice, Wistar rats and sheep vaccinated with Sj14/GST, compared to non-vaccinated control groups. Worm reductions of 48.8% and 49.0% were recorded in Balb/c mice immunized with Sj14/His, compared to non-vaccinated and BCG-vaccinated groups, respectively. These results indicate that rSj14FABP is a promising candidate vaccine for schistosomiasis japonica, particularly as in the rat and sheep vaccination experiments, no adjuvant was used.
Liu
J M
JM
Shanghai Institute of Animal Parasitology, Ministry of Agriculture, Shanghai, PR China. jimyliu48@hotmail.com
Cai
X Z
XZ
Lin
J J
JJ
Fu
Z Q
ZQ
Yang
G Z
GZ
Shi
F H
FH
Cai
Y M
YM
Shen
W
W
Taylor
M G
MG
Wu
X F
XF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D050556:Fatty Acid-Binding Proteins; D005243:Feces / Q000469:parasitology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010270:Parasite Egg Count / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000662:veterinary; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012549:Schistosoma japonicum / Q000235:genetics* / Q000276:immunology*; D012554:Schistosomiasis japonica / Q000276:immunology* / Q000517:prevention & control*; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology / Q000469:parasitology*; D014611:Vaccination / Q000379:methods / Q000662:veterinary*
D002352:Carrier Proteins; D016054:DNA, Protozoan; D050556:Fatty Acid-Binding Proteins; D011994:Recombinant Proteins