›› 2013, Vol. 31 ›› Issue (6): 13-447-449.

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Cloning and Bioinformatics Analysis of Rhoptry Protein 11 of Toxoplasma gondii

ZHANG Xiao-lei,ZHANG Jin-shun,JIA Xiao-hui,XU Yun-peng,ZHANG Ying,WANG Chun-miao,WANG Yan,LU Zhi-min,ZHAO Jian-ling,JIA Tian-jun   

  1. Institute of Pathogen Microbes and Immunology,Hebei North University,Zhangjiakou 075000,China
  • Online:2013-12-30 Published:2014-02-28

Abstract: Total RNA was extracted from tachyzoites of RH strain of Toxoplasma gondii. The open reading frame of ROP11 gene was amplified by using a pair of specific primers designed according to the coding sequence of ROP11 gene (Accession No. DQ077905). The RT-PCR product was digested by restriction enzyme EcoRⅠ and NotⅠ, and then ligated into a pGEX-6P-2 vector. The recombinant plasmid was transferred into E. coli XL-Blue. The positive clones was selected by colony PCR, and confirmed by the double restriction enzyme digestion and sequencing. The RT-PCR product was 1 548 bp. The recombinant plasmid was confirmed by colony PCR and double restriction enzyme digestion. Sequencing results showed that the obtained ROP11 gene was 1 548 bp (Accession No. KC456639). There was a high sequence consistency (99%) between the obtained ROP11 gene sequence and the Toxoplasma ROP11 gene from GenBank. Bioinformatics analysis showed that the ROP11 protein (Mr 57 020) consisted of the signal peptide (amino acids 1-26), 12 conservative domains, a serine/threonine protein kinase catalytic domain (amino acids 170-511), and two potential N-glycosylation sites.

Key words: Toxoplasma gondii, Rhoptry protein 11, Prokaryotic expression, Bioinformatics analysis