ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2014, Vol. 45 ›› Issue (6): 960-966.doi: 10.11843/j.issn.0366-6964.2014.06.014

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Generation and Identification of a Histidine Tagged Recombinant Foot and Mouth Disease Virus

YANG Bo1,2,YANG Fan2,WANG Song-hao2,ZHANG Yan2,YUE Cheng 1*, ZHENG Hai-xue2*,YIN Hong2*   

  1. (1.College of Veterinary Medicine,Xinjiang Agricultural University, Urumqi 830052, China; 2.State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory,Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China)
  • Received:2013-12-19 Online:2014-06-23 Published:2014-06-23

Abstract:

To identify a feasible insertion site in the foot-and-mouth disease virus (FMDV) and obtain a tagged recombinant virus,in this study,we have used reverse genetics to introduce a histidine (His)tag into the ninth site in the downstream of RGD motif in FMDV and obtain His tagged full-length-cDNA clone, named prAsia1-9His.We transfected the prAsia1-9His into BHK-21 cells and typical cytopathic effect (CPE) were found in the transfacted BHK-21 cells.Then,RT-PCR,indirect immunofluorescence assay and sequence analysis were performed,the results showed that the recombinant virus named as rAsia1-9His was successfully rescued and the His tag was stably maintained.Finally,we compared rAsia1-9His with the parental virus (named rAsia1) for pathogenicity to BHK-21 cells and suckling mouse,the results showed that they had similarity in the biological characteristics.The successful rescue of tagged FMDV lays a foundation for further study of molecular pathogenesis and marker vaccine of FMDV.

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