Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (10): 2175-2182.doi: 10.3864/j.issn.0578-1752.2013.10.024

• RESEARCH NOTES • Previous Articles    

Cloning and Expressing Glideosome Associated Protein Genes of Babesia motasi and Preparation of Their Polyclonal Antibodies

 WANG  Jin-Ming, LIU  Ai-Hong, REN  Qiao-Yun, MA  Mi-Ling, LIU  Zhi-Jie, LI  An-Yan, LI  You-Quan, ZHAO  Shuai-Yang, ZHANG  Hao-Hao, YIN  Hong, LUO  Jian-Xun, GUAN  Gui-Quan   

  1. State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Lanzhou 730046
  • Received:2012-08-10 Online:2013-05-15 Published:2013-04-08

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Abstract: 【Objective】To prepare polyclonal antibodies of component genes from glideosome, gap45, gap50, myo A, and mlc were cloned and expressed in prokaryotic system. 【Method】 Rapid amplification of cDNA end (RACE) was used to amplify the full lengths of gap45, gap50, myo A, and mlc. The partial or complete open reading frames (ORF) of those genes were further inserted into prokaryotic expression vector and then transformed into BL21 (DE3) pLys to express by inducing with IPTG. The purified fusion proteins, with MagneGST™ Protein Purification System, were injected into rabbits to produce polyclonal antibodies. Specificities and titers of the polyclonal antibodies were determined by ELISA assay. 【Result】The full-length cDNAs of gap45, gap50, myo A, and mlc, were 664, 1 391, 2 538, 773 bp and open reading frames (ORF) were 567, 1 194, 2 514, and 606 bp in size. The polyclonal antibodies prepared had good specificities and the titers were more than 1:25 600.【Conclusion】In the present study, the full length of four component genes of B. motasi glideosome were amplified and their polyclonal antibodies were obtained, which will provide important biological materials for screening and identifying candidate antigens of vaccination and diagnosis, drug targets as well studying invasion mechanism of Babesia in the future.

Key words: Babesia , RACE , glideosome , clone , expression , polyclonal antibody

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