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猪瘟病毒3’非编码区的多态性及其二级结构分析 预览

Heterogeneity and Secondary Structure Analysis of 3' Untranslated Region in Attenuated Classical Swine Fever Viruses
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摘要 【目的]分析CSFV3'-UTR一级序列和二级结构,为研究其对病毒复制、增殖和毒力的关系奠定良好的基础。【方法】利用RT-PCR技术扩增CSFVThiverval株、HCLV株和Shimen株的3'-UTR并测序,使用DNAstar、Sequencher和RNAstructurc软件进行CSFV3'-UTR基因组序列的比对分析和二级结构模拟。【结果】Thiverval株3'-UTR最长可达259个碱基,与母源株Alfort相比,含有32nt的插入,最短只有233个碱基,含有6nt的插入。HCLV株最长有244个碱基,含有17nt的插入;最短有233个碱基,含有8nt的插入。同时,疫苗株的插入片段导致3'-UTR空间构型和自由能发生改变,自由能随着插入片段的增加而升高。【结论】(1)CSFV3'-UTR存在两个高度变异的区域。(2)CSFV疫苗株Thiverval株和HCLV株3'-UTR同一样品不同克隆株中插入片段呈现明显的不均一。(3)疫苗株的插入片段影响3'-UTR二级结构的稳定性,可能是导致疫苗株毒力减弱的原因之一。 [ Objective] The 3'-UTR of CSFV was sequenced and analyzed, for the understanding of the influence of 3'-UTR on replication, multiplication, and virulence of virus. [Method] The 3'-UTR of CSFV Thiverval, HCLV and Shimen strains were amplified and sequenced using RT-PCR method. The alignment was carried out using DNAstar and Sequencher software. The RNA secondary structures of different 3'-UTR were predicted using RNAstructure software. [ Result] The longest 3'-UTR of Thiverval strain was 259 base pairs (bp) with a 32 nt insertion. The shortest 3'-UTR had only 233 bp with a 6 nt insertion. The longest 3'-UTR of HCLV strain was 244 bp with a 17 nt insertion and the shortest 3'-UTR was 235 bp with a 8 nt insertion. At the same time, to maintain the similar 3'-UTR structure, the free energy of vaccine strain is higher than that of virulent strain, and the free energy in 3'-UTR of vaccine strain is rising with length of insert. [Conclusion] The 3'-UTR of CSFV vaccine strains have two variable regions. The insertion of attenuated Thiverval and HCLV strains 3'-UTR represent distinct heterogeneity in different clones of the same sample. The insertion of vaccine strains 3'-UTR could affect the stability of the secondary structure, which might be a reason for the attenuated virulence.
作者 范运峰 赵启祖 赵耘 邹兴启 张仲秋 王琴 宁宜宝 FAN Yun-feng, ZHAO Qi-zu, ZHAO Yun, ZOU Xing-qi, ZHANG Zhong-qiu, WANG Qin, NING Yi-bao(1China Institute of Veterinary Drug Control, Beijing 100081; 2China Animal Disease Control Center, Beijing 100125)
出处 《中国农业科学》 CAS CSCD 北大核心 2010年第4期828-834,共7页 Scientia Agricultura Sinica
基金 国家自然科学基金资助项目(30571377)和国家“863”计划项目(2006AA10A204)
关键词 猪瘟病毒 3'-UTR 不均一性 二级结构分析 classical swine fever virus 3'-UTR heterogeneity RNA secondary structure
作者简介 范运峰,博士。Tel-010—59194743 E—mail:fanyunfeng2000@yahoo.com.cn。 通信作者赵启祖,研究员。Tel:010-62103677;E-mail:zhaoqizu@ivdc.gov.cn
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