title: Construction and characterization of an infectious clone of coxsackievirus A16.
abstract: BACKGROUND
  Coxsackievirus A16 (CVA16) is a member of the Enterovirus genus of the Picornaviridae family and it is a major etiological agent of hand, foot, and mouth disease (HFMD), which is a common illness affecting children. CVA16 possesses a single-stranded positive-sense RNA genome containing approximately 7410 bases. Current understanding of the replication, structure and virulence determinants of CVA16 is very limited, partly due to difficulties in directly manipulating its RNA genome.
  
  
  RESULTS
  Two overlapping cDNA fragments were amplified by RT-PCR from the genome of the shzh05-1 strain of CVA16, encompassing the nucleotide regions 1-4392 and 4381-7410, respectively. These two fragments were then joined via a native XbaI site to yield a full-length cDNA. A T7 promoter and poly(A) tail were added to the 5' and 3' ends, respectively, forming a full CVA16 cDNA clone. Transfection of RD cells in vitro with RNA transcribed directly from the cDNA clone allowed the recovery of infectious virus in culture. The CVA16 virus recovered from these cultures was functionally and genetically identical to its parent strain.
  
  
  CONCLUSIONS
  We report the first construction and characterization of an infectious cDNA clone of CVA16. The availability of this infectious clone will greatly enhance future virological investigations and vaccine development for CVA16.
authors.affiliation: Key Laboratory of Molecular Virology & Immunology, Institute Pasteur of Shanghai, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Liu
authors.forename: Qingwei
authors.initials: Q
authors.lastname: Liu
authors.forename: Yicun
authors.initials: Y
authors.lastname: Cai
authors.forename: Qibin
authors.initials: Q
authors.lastname: Leng
authors.forename: Zhong
authors.initials: Z
authors.lastname: Huang
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods*; D003588:Cytopathogenic Effect, Viral; D018076:DNA, Complementary / Q000235:genetics* / Q000378:metabolism; D004770:Enterovirus / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity*; D016679:Genome, Viral; D006232:Hand, Foot and Mouth Disease / Q000821:virology; D006801:Humans; D012367:RNA, Viral / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014709:Vero Cells; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism*; D014774:Virulence
chemical_list: D018076:DNA, Complementary; D012367:RNA, Viral; D015678:Viral Structural Proteins