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[Induced expression of the variable region of AMA-1 from Plasmodium falciparum]
1Department of Etiological Biology, Faculty of Basic Medicine Sciences, Second Military Medical University, Shanghai 200433.
Objective:
To express the variable region of AMA-1 gene from Plasmodium falciparum in Escherichia coli.
Methods:
Genomic DNA of FCC1/HN was used as template and the variable region of AMA-1 gene was amplified by polymerase chain reaction(PCR). The PCR products were digested by endonuclease BamH I and Hind III, cloned into pBlu2KSP. The nucleotide sequences of the variable region of AMA-1 gene were determined by sequencing. The AMA-1 gene fragment was subcloned into plasmid pQE, expressed in E. coli and induced by IPTG. The fusion product as identified by SDS-PAGE gel electrophoresis and Western blotting were proceeded with anti-AMA-1 sera from rabbit.
Results:
The size of the variable region of AMA-1 gene from FCC1/HN was 506 bp and encoded 168 amino acids. On SDS-PAGE gel dyed with Coomassie brilliant blue R250, no specific protein band can be discerned, but Western blotting proceeded with anti-AMA-1 sera from rabbit demonstrated that the specific protein band was about 23.0 kDa.
Conclusion:
The variable region of AMA-1 gene from FCC1/HN was able to be expressed in E. coli and analysis of Western blotting demonstrated that the AMA-1 fussion protein contained specific antigenic epitopes.