›› 2007, Vol. 25 ›› Issue (1): 8-40.

• 实验研究 • Previous Articles     Next Articles

Giardia canis Virus Transfection Vector-Mediated Expression of Green Fluorescent Protein in the Parasite

CHEN Li-feng1;LI Jian-hua1;LIU Quan2;ZHAO Yue-ping1;CAO Li-li1;ZHANG Xi-chen1   

  1. 1 College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China; 2 Hebei Normal University of Science ＆ Technology, Qinhuangdao 066000, China; 3 Veterinary Institute, Academy of Military Medical Sciences, Changchun 130062, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2007-02-28 Published:2007-02-28
  • Contact: ZHANG Xi-chen

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Abstract: Objective To construct Giardia canis virus (GCV) transfection vector. Methods According to transcriptional start site, replication origin and packaging site of GCV genome (DQ238861), a system was developed for the expression of a foreign gene in this organism by flanking the green fluorescent protein (GFP) gene with the fragments of GCV positive-strand RNA. The transcript of the construct was synthesized in vitro with T7 RNA polymerase and used to transfect GCV-infected trophozoites by electroporation. Results The recombinant plasmid pGCV634/GFP/GCV2174 was constructed. The expression of green fluorescent protein mediated by GCV transfection vector in Giardia canis peaked at 1 d after electroporation(A490=1.8), and slowly decreased until 14 d post-transfection. Conclusion The engineered GCV vector can be successfully used to introduce and efficiently express a heterologous gene in the eukaryotic microorganism.

Key words: Giardia canis virus, Transfer vector, Green fluorescent protein