ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2015, Vol. 46 ›› Issue (4): 608-614.doi: 10.11843/j.issn.0366-6964.2015.04.014

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The Mutations of 3′-U3 Region Make No Difference to ALV’s Replication Capacity in vitro

GAO Yan-ni,GAO Qi,LI Xiao-fei,YUN Bing-ling,QI Xiao-le,WANG Yong-qiang,LIU Chang-jun,CUI Hong-yu,ZHANG Yan-ping,GAO Hong-lei,WANG Xiao-mei,GAO Yu-long*   

  1. (State Key Laboratory of Veterinary Biotechnology/Division of Avian Immunosuppressive Diseases,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China)
  • Received:2014-08-14 Online:2015-04-23 Published:2015-04-23

Abstract:

In order to study the influence of the 3′-U3 region with regular mutations of epidemic J subgroup avian leukosis virus (ALV-J) strains isolated from egg-type flocks on its replication capacity in vitro,a full-length cDNA clone was constructed based on ALV-J prototype HPRS-103 strain infectious clone via replacing its 3′-U3 region by the corresponding part of egg-type isolate SD09DP04.Both the constructed chimeric infectious clone and the HPRS-103 infectious clone were transfected into DF1 cells by lipofectamine 2000,respectively.The two infectious clones were passaged to another generation.And the harvested viruses were respectively confirmed by indirect immunofluorescence (IFA),ALV-antigen test kit and reverse transcriptase activity test kit.Analyzing the 3′-U3 region’s promoter activity or enhancer activity and TCID50 growth curves showed that such U3 region with regular mutations had no evident effect on the virus’s replication capacity in vitro.The successful rescue of the chimeric virus plays the basic role in further exploration of the functions and the significance of the regular mutations in the 3′-U3 region in the epidemic ALV-J strains in recent years.

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