[Cloning, expression and antigenic analysis of merozoite surface protein MSPDBL2-DBL2 domain from Plasmodium falciparum].
OBJECTIVE
To clone and express the DBL domain of Plasmodium falciparum merozoite surface protein MSPDBL2(DBL2), and investigate its antigenicity.


METHODS
The DBL2 fragment was amplified by PCR and cloned into pET28a vector. The recombinant pET28a-DBL2 plasmid was transformed into E. coli BL21 (DE3) and protein expression was induced by IPTG. The expressed product was purified by Ni-NTA affinity chromatography, and analyzed by SDS-PAGE and Western blotting.


RESULTS
DBL2 gene fragment of Plasmodium falciparum merozoite surface protein MSPDBL2 (950 bp) was obtained by PCR. The recombinant pET28a-DBL2 plasmid was identified by PCR, double enzyme digestion, and DNA sequencing. The recombinant DBL2 protein was expressed in an inclusion body form with Mr 340,000 after being induced with IPTG. Moreover, the purified recombinant DBL2 protein was recognized by sera from patients with falciparum malaria.


CONCLUSIONS
The recombinant pET28a-DBL2 plasmid has been constructed. The purified rDBL2 protein shows adequate antigenicity.
Meng-jia
MJ
Yang
Su-rong
SR
Wang
Hong-xiang
HX
Zhuge
Jun-hu
JH
Chen
D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology*; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology*; D052940:Merozoites / Q000276:immunology*; D010963:Plasmodium falciparum / Q000276:immunology*; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins
D000953:Antigens, Protozoan; D008565:Membrane Proteins; D011994:Recombinant Proteins