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Published on: August 22, 2019
[Cloning, prokaryotic expression and immunoreactivity evaluation of Angiostrongylus cantonensis galectin]
Li Hao1, Kun Wu, Xiao-guang Chen
1Department of Pathogenic Biology, College of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China. haoli80@126.com
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.
Conclusion:
The success in cloning and identification, the recombinant AcGAL may provide basis for further diagnostic study of angiostrongyliasis.
