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含74个腺嘌呤poly(A)尾的猪水泡病病毒HK/70株
3′非编码区的克隆及其特征分析
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郑海学1，2,刘湘涛1，2,尚佑军1,吴锦艳1,冯霞1,白兴文1
田宏1，3,孙世琪1，2,尹双辉1,郭建宏1，2，刘在新1，2,谢庆阁1，2
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（1. 中国农业科学院 兰州兽医研究所 农业部畜禽病毒学重点实验室，甘肃 兰州730046；
2. 中国农业科学院 研究生院，北京 100081; 3. 西北农林科技大学，陕西 杨凌712100）
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摘要：
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应用3′RACE法降落式扩增并克隆了猪水泡病病毒（SVDV）HK/70株基因组3′端的真实序列。测序结果表明，所扩增的目的基因片段长4 200 nt，包括非结构蛋白编码区（P2和P3）、3′端非编码区和poly(A)尾，其中3′NTR位于终止密码子TAA之后，长99 nt，poly(A)至少含有74个腺嘌呤碱基。经与参考毒株进行序列比较，结果显示, HK/70株与J1′73、H/3′76和AUG/22/90株的核苷酸同源性较高，为99.0%（仅第65位碱基不同），而与NET/1/92参考毒株的核苷酸同源性较低，为83.3%，与人柯萨奇病毒B5 UK/54株(UK/54CB5)的同源性为75.7%。同时对3′NTR的二级结构进行分析，并与参考毒株和CB5毒株的二级结构进行了比较。结果表明，它们具有协同变异性，有共同的祖先，显示出3′NTR存在支持其功能所必需的一些结构域。 |
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关键词：
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猪水泡病病毒； 3′NTR； poly(A)； 二级结构； 柯萨奇病毒B5 |
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中图分类号：S 852.659.6 文献标识码：
A 文章编号：1673-4696(2006)01-0007-06 |
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Molecular cloning and characterization of 3′ nontranslated
region of swine vesicular disease virus HK/70 strain
with 74 adenines poly(A) tail |
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ZHENG Haixue1，2, LIU
Xiangtao1，2, SHANG Youjun1, WU
Jinyan1, FENG Xia1,BAI
Xingwen1,
TIAN Hong1，3, SUN Shiqi1，2, YIN
Shuanghui1, GUO Jianhong1,2, LIU
Zaixin1，2,XIE Qingge1，2 |
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(1.Key Laboratory of Animal Virology, Ministry of Agriculture/ Lanzhou Veterinary Research Institute, Chinese
Academy of Agricultural Sciences, Lanzhou 730046, China; 2. Graduate School,
Chinese Academy of Agricultural Sciences, Beijing 100081, China; 3.Northwest SciTech
University of Agriculture and Forestry, Yangling 712100, China) |
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Abstract: The authentic 3′end sequence, including P1, P2, 3′NTR, and poly (A) tail, was amplified by 3′RACE, and the results showed that 3′NTR was 99 nt in length, and the poly(A) tail had at least had 74 adenines. Compared with other reference strains, HK/70 strain shared higher nucleotide acid identity with J1′73, H/3′76 and AUG/22/90 strains(99.0%) than with NET/2/92 and UK/54CB5(83.3% and 75.7%), The secondary structure of HK/70 strain was predicted and analyzed by RNAdraw software, and results showed that there were some special domains in the 3′NTR, which was necessary for its biological function, and there are also some covariable positions supporting the hypothesis which SVDV most likely arose from a single common ancestor. SVDV may be a recently evolved genetic subspecies of CB5. |
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Key words: swine vesicular disease virus; 3′NTR; poly(A); secondary structure; Coxsackiev B5 |
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