ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (5): 706-711.doi:

• 预防兽医 • Previous Articles     Next Articles

Construction of an Infectious Full-length cDNA Clone of Foot-and-mouth Disease Virus Asia1/JS/China/2005 Strain

LI Ping-hua1, BAI Xing-wen1, LU Zeng-jun1, SUN Pu1, GUO Jian-hong1,
CAO Wei-jun2, LIU Xiang-tao1, YIN Hong1*, LIU Zai-xin1*   

  1. 1. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, National FootandMouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Lanzhou 730046, China；2. Gansu Agricultural University, Lanzhou 730070，China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-05-25 Published:2009-05-25

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Abstract: Three overlapping cDNA fragments covering the approximate FMDV full genome (7.5kb）of the 3′ end viral genome were amplified using long RTPCR and were cloned into pBluescriptSK+ vector with unique restriction sites, respectively. A fragment (about 0.7 kb ) including 15 C of the 5′ end viral genome was amplified by over-lap PCR and was cloned into pGEM-T vector. All positive clones were assembled into a low copy number vector pCDNA31/Zeo（+）which was removed T7 promoter in the vector sequence and constructed FMDV Asia1/JS/China/2005 full-length cDNA clone. RNA was synthesized in vitro using T7 polymerase, the infective virus was obtained by transfecting the RNA into BHK-21 cells. The rescued virus was identified by the RT-PCR, indirect immunofluorescence, electron microscope and sulk mice pathogenicity, the results showed infectious FMDV was rescued successfully. The full-length infectious cDNA clone will lays the basis for elucidating the mechanism of pathogenesis of FMDV and developing novel vaccines against FMD.