[Cloning, prokaryotic expression and immunoreactivity evaluation of Angiostrongylus cantonensis galectin].
OBJECTIVE
To construct the recombinant plasmid for Angiostrongylus cantonensis (AC) galectin (GAL) cDNA and analyze the immunological activity of the recombinant protein.


METHODS
AcGAL 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.


RESULTS
For 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.


CONCLUSIONS
The success in cloning and identification, the recombinant AcGAL may provide basis for further diagnostic study of angiostrongyliasis.
Hao
Li
L
Department of Pathogenic Biology, College of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China. haoli80@126.com
Wu
Kun
K
Chen
Xiao-guang
XG
Wang
Qiong
Q
D017185:Angiostrongylus cantonensis / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D037161:Galectins / Q000235:genetics / Q000378:metabolism*; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000378:metabolism*; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D037161:Galectins; D015801:Helminth Proteins; D011994:Recombinant Proteins