title: A simple and novel method for GII norovirus genome clone with generic primers.
abstract: OBJECTIVE
  This study aims to establish a novel method for cloning GII norovirus genome using generic primers rationally designed based on multiple alignments of 96 GII norovirus genome sequences.
  
  
  RESULTS
  Based on conservative analysis of 96 GII norovirus genome sequences available in GenBank, three fragments encompassing the full-length genome were rationally designed. Fragments A, B and C were amplified by primers N1F/N2819R, N2689F/COG2R and COG2F/adaptor, respectively. Meanwhile, the sensitivity of the novel primers was evaluated, which could achieve 10(1) RTPCRU, as determined by the common detection primer pair JV12/JV13. The availability of the novel protocol was verified by sequencing two norovirus strains with different genotypes.
  
  
  CONCLUSIONS
  Primers for GII norovirus genome clone were rationally designed, and a novel GII genome clone method was established.
  
  
  CONCLUSIONS
  The three-fragment cloning method can be used as a universal tool to collect information on the genome of norovirus strains for future evolution and antivirus studies.
authors: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, Guangdong, China.
authors: L
authors: L
authors: Xue
authors: Q
authors: Q
authors: Wu
authors: X
authors: X
authors: Kou
authors: J
authors: J
authors: Zhang
authors: W
authors: W
authors: Guo
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D017931:DNA Primers / Q000737:chemistry*; D016679:Genome, Viral; D005838:Genotype; D029322:Norovirus / Q000235:genetics*; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers
keywords: genome clone; norovirus; phylogenetic analysis; rational design; reverse transcriptase-polymerase chain reaction