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

Authors

  • L. Xue,

    1. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, Guangdong, China
    2. Guangdong Institute of Microbiology, State Key Laboratory of Applied Microbiology (Ministry-Guangdong Province Jointly Breeding Base), South China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Open Laboratory of Applied Microbiology, Guangzhou, Guangdong, China
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  • Q. Wu,

    Corresponding author
    1. Guangdong Institute of Microbiology, State Key Laboratory of Applied Microbiology (Ministry-Guangdong Province Jointly Breeding Base), South China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Open Laboratory of Applied Microbiology, Guangzhou, Guangdong, China
    • School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, Guangdong, China
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  • X. Kou,

    1. Guangdong Institute of Microbiology, State Key Laboratory of Applied Microbiology (Ministry-Guangdong Province Jointly Breeding Base), South China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Open Laboratory of Applied Microbiology, Guangzhou, Guangdong, China
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  • J. Zhang,

    1. Guangdong Institute of Microbiology, State Key Laboratory of Applied Microbiology (Ministry-Guangdong Province Jointly Breeding Base), South China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Open Laboratory of Applied Microbiology, Guangzhou, Guangdong, China
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  • W. Guo

    1. Guangdong Institute of Microbiology, State Key Laboratory of Applied Microbiology (Ministry-Guangdong Province Jointly Breeding Base), South China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Open Laboratory of Applied Microbiology, Guangzhou, Guangdong, China
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Correspondence

Qingping Wu, Guangdong Institute of Microbiology, State Key Laboratory of Applied Microbiology (Ministry-Guangdong Province Jointly Breeding Base), South China Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Open Laboratory of Applied Microbiology, No. 100, Xianlie Zhong Road, Guangzhou 510070, China.

E-mail: wuqp203@yahoo.com.cn

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/COG2R and COG2F/adaptor, respectively. Meanwhile, the sensitivity of the novel primers was evaluated, which could achieve 101 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.

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