[Gene cloning, construction and expression of single-chain Fv (scFv) against the membrane protein of Schistosoma japonicum].
OBJECTIVE
To construct single chain antibody specific to membrane protein of Schistosoma japonicum by genetic engineering technique.


METHODS
The VH (heavy-chain variable region) and VL(light-chain variable region) genes were amplified by PCR from the genomic DNA of NP11-4 cell line, and sequenced by Sanger's method. The ScFv was constructed in pTHA90 vector using VH and VL genes, then expressed by IPTG.


RESULTS
The VH and VL genes were obtained through PCR. The DNA sequences showed that VH and VL were new variable region genes of antibody. They were registered by GenBank. A ScFv gene with (Gly4Ser) 3 intralinker in the pTHA90 vector was successfully constructed. The ScFv was expressed as thioredoxin-fused proteins about 36.2 kDa.


CONCLUSIONS
A specific ScFv against the membrane protein of Schistosoma japonicum was constructed and expressed.
Yu
X C
XC
Institute of Genetics, Chinese Academy of Sciences, Beijing 100101.
Jiang
X
X
Huang
H M
HM
Zhang
Z
Z
Lin
Q
Q
Guan
X H
XH
Huang
H L
HL
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D005818:Genetic Engineering; D015801:Helminth Proteins / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis* / Q000235:genetics; D008565:Membrane Proteins / Q000276:immunology*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D012549:Schistosoma japonicum / Q000276:immunology*; D017422:Sequence Analysis, DNA
D000909:Antibodies, Helminth; D015801:Helminth Proteins; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins