关键词: 间日疟原虫； 疟疾； Duffy血型结合蛋白； 原核表达； 疫苗； 中国

Abstract:

Objective To clone a Plasmodium vivax Duffy binding protein critical functional region II（PvDBPII）gene of central China isolate，and to express and identify the recombinant PvDBPII protein in vitro. Methods PCR was performed to am? plify PvDBPII from P. vivax DNA of a central China isolate and the PCR product was inserted into pET28a （+）vector. pET28a? PvDBPII recombinant plasmid was constructed and transformed into E. coli host BL21 （DE3+） . IPTG was used to induce the recom? binant PvDBPII 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 blot. Results The PCR product of PvDBPII gene was about 1.1 kb，meeting the expectation of predicted fragment size. The recombinant pET28a?PvDBPII plasmid was verified by sequencing that the insertion was correct both in direction and in frame，but with 4 non?synonymous mutations compared to reference P. vivax strain Sal?I. SDS? PAGE，and Western blot analysis showed that the recombinant PvDBPII protein was about 44 kDa，and could be recognized by pooled sera from vivax malaria patients. Conclusion The PvDBPII gene of central China isolate is successfully cloned，and re? combinant PvDBPII is expressed， thereby providing opportunity for further study on PvDBPII.

Key words: Plasmodium vivax；Malaria；PvDBPII； Prokaryotic expression； Vaccine； China