关键词: 恶性疟原虫, AMA-1基因变异区, PCR, 克隆, 大肠杆菌, 基因表达

Abstract: 　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 Bam HⅠ and Hin dⅢ, 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.

Key words: Plasmodium falciparum, variable region of AMA-1, PCR, cloning, Escherichia coli, gene expression