JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2017, Vol. 55 ›› Issue (3): 64-69.doi: 10.6040/j.issn.1671-7554.0.2016.104

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Bioinformatics analysis and construction of an eukaryocyte vector of ROP19 protein in Toxoplasma gondii

ZHOU Jian1, ZHAO Hanting1, LÜ Gang2, WANG Lin3, LI Qihang1, ZHU Meiyan1, WANG Zhilin1, HE Shenyi2   

  1. 1. School of Medicine, Shandong Univeristy, Jinan 250012, Shandong, China;
    2. Department of Parasitology, School of Basic Medical Sciences, Shandong Univeristy, Jinan 250012, Shandong, China;
    3. Department of Nerve Electrophysiology, Jinan Childrens Hospital, Jinan 250022, Shandong, China
  • Received:2016-02-02 Online:2017-03-10 Published:2017-03-10

Abstract: Objective To construct and express the eukaryotic expression vector of ROP19 protein of Toxoplasma gondii PRU strain. Methods The physical and chemical characteristics of ROP19 protein were analyzed, and the B cell epitopes and T cell epitopes of ROP19 were compared with SAG1 using bioinformatics. The recombinant eukaryotic expression plasmid pROP19 was constructed by the molecular cloning technology. After being identified by PCR, restriction enzyme cleavage and sequencing, pROP19 was transfected into the HEK293T cells. The cells from different groups(control, pEGFP-C1 and pROP19)were respectively detected with fluorescence microscope under blue laser. The ROP19 protein was detected by Western blotting. Results The ROP19 protein was mainly located in the membrane and had better antigenic index than SAG1. The plasmid was constructed successfully. Western blotting showed that the expressed proteins could be recognized by anti-STAg mouse sera. Conclusion The eukaryotic expression vector pROP19 is constructed and expressed in eukaryotic cells.

Key words: Eukaryotic expression, ROP19 protein, Toxoplasma gondii, Bioinformatics

CLC Number: 

  • R382.5
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