title: [Reconstituting turkey herpesvirus with bacterial artificial chromosome clones].
abstract: Herpesvirus of turkey (HVT) is an alpherpesvirus and widely used as a live vaccine against Marek's disease (MD) because of its antigenic relationship with Marek's disease virus (MDV).
  
  
  OBJECTIVE
  The aim of this study was to construct Herpesvirus of turkey Fc126 strain as an infectious bacterial artificial chromosome (BAC).
  
  
  METHODS
  Using the selection marker Eco-gpt (Xanthine-guanine phosphoribosyl transferase)(1.3 kb) and BAC vector pBeloBAC11(7.4 kb), we constructed the transfer plasmid pGAB-gpt-BAC11. Then, the transfer plasmid and HVT-infected cells' total DNA were cotransfected into primary chicken embryo fibroblasts (CEFs). After six rounds of selection in medium containing mycophenolic acid, xanthine and hypoxanthine, we obtained purified recombinant viruses. Genomic DNA was extracted and electroporated into Escherichia coli DH10B competent cells. BAC clones were identified by restriction enzyme digestion and PCR analysis, and then tested for infectivity after transfection into CEFs using calcium phosphate.
  
  
  RESULTS
  We obtained 25 BAC clones, and reconstituted recombinant viruses by transfection of HVT-BAC6 DNA, HVT-BAC8 DNA and HVT-BAC10 DNA into CEFs respectively.
  
  
  CONCLUSIONS
  In this study, we cloned the complete genome of HVT Fc126 strain as an infectious bacterial artificial chromosome.
authors: Division of Avian Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute the Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors: Desong
authors: D
authors: Lan
authors: Xingming
authors: X
authors: Shi
authors: Yunfeng
authors: Y
authors: Wang
authors: Hongyu
authors: H
authors: Cui
authors: Changjun
authors: C
authors: Liu
authors: Mei
authors: M
authors: Wang
authors: Wenwei
authors: W
authors: Hu
authors: Guangzhi
authors: G
authors: Tong
mesh_terms: D000818:Animals; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D026002:Herpesvirus 1, Meleagrid / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D014162:Transfection; D014779:Virus Replication
chemical_list: D004274:DNA, Recombinant; D004279:DNA, Viral