Cloning, prokaryotic expression and immunoreactivity evaluation of Angiostrongylus cantonensis galectin.
- Author:
Li HAO
1
;
Kun WU
;
Xiao-guang CHEN
;
Qiong WANG
Author Information
- Publication Type:Journal Article
- MeSH: Angiostrongylus cantonensis; genetics; metabolism; Animals; Blotting, Western; Cloning, Molecular; DNA, Complementary; chemistry; genetics; Electrophoresis, Polyacrylamide Gel; Escherichia coli; genetics; Galectins; genetics; metabolism; Helminth Proteins; genetics; metabolism; Molecular Sequence Data; Recombinant Proteins; metabolism; Sequence Analysis, DNA
- From: Journal of Southern Medical University 2007;27(5):584-587
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the recombinant plasmid for Angiostrongylus cantonensis (AC) galectin (GAL) cDNA and analyze the immunological activity of the recombinant protein.
METHODSAcGAL cDNA was screened from the cDNA library and amplified by PCR. The amplified fragment was subcloned into the expression vector pET32a(+) and expressed in E.coli. The inclusion body was washed, degenerated, refolded by dialysis, and condensed for SDS-PAGE and Western blot analysis of the protein.
RESULTSFor the first time the full-length cDNA of AcGAL was cloned (GenBank GeneID: DQ384534). Restriction enzyme digestion indicated that the recombinant plasmid pET32a(+)-AcGAL was successfully constructed. SDS-PAGE analysis confirmed high expression of the recombinant protein AcGAL in E.coil in the form of inclusion bodies, which possessed good immunoreactivity as shown by Western blot analysis.
CONCLUSIONThe success in cloning and identification, the recombinant AcGAL may provide basis for further diagnostic study of angiostrongyliasis.
