A simple and novel method for GII norovirus genome clone with generic primers

作者：Xue, L.; Wu, Q.^*; Kou, X.; Zhang, J.; Guo, W.

来源：Journal of Applied Microbiology, 2013, 115(2): 613-620.

DOI：10.1111/jam.12244

• genome clone
• norovirus
• phylogenetic analysis
• rational design
• reverse transcriptase-polymerase chain reaction

Aims: 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.
Methods and 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(I) 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.
Significance and Impact of the Study: 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.

• 出版日期2013-8
• 单位广东省微生物研究所; 华南理工大学