[Expression of Schistosoma japonicun fatty acid binding protein gene in silkworm cells and larvae].
OBJECTIVE
To express the fatty acid binding protein (Sj14FABP) gene of Schistosoma japonicun in the silkworm cells and larvae.


METHODS
A 600 bp DNA fragment containing Sj14FABP gene was cloned into baculovirus transfer vector of pBacPAK His1 to construct recombinant transfer vector Sj14-pBac PAK His1. Coinfection was accomplished with this vector and Bombyx mori nuclear polyhedrosis virus (BmNPV) DNA in BmN cells. The recombinant virus of Bm-Sj14 was screened using dot-blotting. The BmN cells and silkworm larvae were infected with Bm-Sj14 to express Sj14FABF gene. Western blotting and ELISA were used to identify the antigenicity of the recombinant protein.


RESULTS
Sj14FABP gene was successfully expressed in the BmN cells and silkworm larvae infected with Bm-Sj14. The product was a 18 kDa fusion protein. The yield in BmN cells was about 100 micrograms/1 x 10(6) cells and 33 micrograms/ml cell supernatant. In silkworm larvae, the product yield was 4 mg/ml haemolymph as well as 4.6 mg/g silkworm tissue. The recombinant protein could be recognized by Western blotting and ELISA using the sera from mice immunized with SWAP.


CONCLUSIONS
Sj14FABP gene has been successfully expressed in BmNPV system and the product has high antigenicity.
Liu
J
J
Shanghai Institute of Animal Parasitology, CAAS, Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai 200232.
Cai
X
X
Lin
J
J
Yang
G
G
Shen
X
X
Fu
Z
Z
Shi
F
F
Shen
W
W
Li
M
M
Yuan
C
C
Li
H
H
Cai
Y
Y
Wu
X
X
D000818:Animals; D012831:Bombyx / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics*; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D015870:Gene Expression; D005822:Genetic Vectors; D007814:Larva / Q000235:genetics; D051379:Mice; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012549:Schistosoma japonicum / Q000235:genetics*
D002352:Carrier Proteins; C082667:Fabp5 protein, mouse; C086936:Fabp7 protein, mouse; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins