1.Cloning,expression and identification of gametocyte specific protein Pfgdv1 of Plasmodium falciparum
Pangpang SU ; Lingwen MENG ; Jiangyan LI ; Zhiyong TAO ; Yong CHEN ; Jichen QIAO ; Xiaoxiao WU ; Haopeng WANG ; Qiang FANG ; Xuemei WANG ; Hui XIA
Chinese Journal of Schistosomiasis Control 2016;(1):34-38
Objective To clone a gametocyte specific protein Pfgdv1 of Plasmodium falciparum,express and identify re?combinant Pfgdv1 protein in vitro. Methods PCR was performed to amplify Pfgdv1 from P. falciparum DNA which was got from the patient who was infected with P. falciparum,and the PCR product was inserted into pET28a(+)vector. pET28a?Pfg?dv1 recombinant plasmid was constructed and transformed into E. coli host BL21(DE3+). IPTG was used to induce the recombi?nant Pfgdv1 protein fused with His tag,and the protein was purified by His?NTA affinity chromatography. The recombinant pro?tein was identified by SDS?PAGE and Western blotting. Results The PCR product of Pfgdv1 gene was about 1.65 kb,meeting the expectation of predicted fragment size. The recombinant protein was about 67 kDa,which could be recognized by His?Tag monoclonal antibody. Conclusion The Pfgdv1 gene of P. falciparum is successfully cloned,and the recombinant Pfgdv1 pro?tein is expressed,thereby providing an opportunity for further study on transmission blocking vaccine.
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