Cloning and Optimized Prokaryotic Expression of a pbmag-1 cDNA Fragment from Plasmodium berghei ANKA
- VernacularTitle:伯氏疟原虫pbmag-1基因片段克隆及原核表达优化
- Author:
Yuhui GAO
;
Heng WANG
- Publication Type:Journal Article
- Keywords:
pbmag-1;
Plasmodium berghei;
Gene cloning;
Prokaryotic expression
- From:
Chinese Journal of Parasitology and Parasitic Diseases
1987;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone and express a novel gene cDNA fragment,pbmag-1,from Plasmodium berghei ANKA strain. Methods The cDNA sequence of pbmag-1 was obtained from the GenBank of P.berghei ANKA genomic databases,with which a pair of primers was designed and RT-PCR was used to get a cDNA fragment of the gene from the parasite. The expanded cDNA 3′ fragment of the gene was obtained by 3′-RACE using the oligo dT primer and a set of specific primers. The intact cDNA 3′ fragment was cloned into a prokaryotic expressional vector and transformed into the BL21-(DE3)-RIL strain of Escherichia coli. The recombinant protein of PbMAg-1 was expressed with an optimized strategy and used to immunize mice. Results The pbmag-1 cDNA fragment obtained was 1 341 bp in length,A/T rich (73%) and with a correct 3′ end sequence. By Western blot,the anti-serum of mice immunized with the recombinant protein of PbMAg-1/GST,which was expressed as inclusion bodies,specifically recognized a band with Mr 64 000 molecule from the protein extracts of P. berghei-infected mouse erythrocytes. Conclusion The pbmag-1 cDNA sequence with intact 3′ has been obtained,which will be used for further study on its role in the immune response of P. berghei infection.
