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A simple and novel method for GII norovirus genome clone with generic primers
Author(s) -
Xue L.,
Wu Q.,
Kou X.,
Zhang J.,
Guo W.
Publication year - 2013
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12244
Subject(s) - christian ministry , china , library science , geography , biology , political science , computer science , archaeology , law
Abstract 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/ COG 2R and COG 2F/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 JV 12/ JV 13. 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.