[Induced expression of the variable region of AMA-1 from Plasmodium falciparum].
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.


CONCLUSIONS
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.
Nie
B Y
BY
Department of Etiological Biology, Faculty of Basic Medicine Sciences, Second Military Medical University, Shanghai 200433.
Zhang
L X
LX
Pan
W Q
WQ
Qian
F
F
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D014644:Genetic Variation; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000235:genetics*; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics
D000953:Antigens, Protozoan; D008565:Membrane Proteins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C090077:apical membrane antigen I, Plasmodium