| 几种PCR方法在克隆鸭肠炎病毒未知基因中的应用 |
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| 引用本文： | 潘华奇,曹瑞兵,陈溥言,刘 磊,王书锦,潘光炎,胡江春. 几种PCR方法在克隆鸭肠炎病毒未知基因中的应用[J]. 微生物学通报, 2009, 36(12): 1842-1848 |
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| 作者姓名： | 潘华奇 曹瑞兵 陈溥言 刘 磊 王书锦 潘光炎 胡江春 |
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| 作者单位： | 中国科学院沈阳应用生态研究所 辽宁 沈阳 110016; 南京农业大学农业部动物疫病诊断与免疫重点开放实验室 江苏 南京 210095; 甘肃农业大学动物医学院 甘肃 兰州 730070;南京农业大学农业部动物疫病诊断与免疫重点开放实验室 江苏 南京 210095;南京农业大学农业部动物疫病诊断与免疫重点开放实验室 江苏 南京 210095;甘肃农业大学动物医学院 甘肃 兰州 730070;中国科学院沈阳应用生态研究所 辽宁 沈阳 110016;甘肃农业大学动物医学院 甘肃 兰州 730070;中国科学院沈阳应用生态研究所 辽宁 沈阳 110016 |
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| 摘 要： | 以DEV基因组DNA为模板, 用简并PCR、改良Targeted gene walking PCR、改良的热不对称交错PCR和Long-PCR, 获得了5350 bp、11083 bp和2905 bp三段DEV未知基因片段, DNA序列分析发现包含9个开放阅读框, 将这些序列提交GenBank分别获得的登录号为: EF554396~EF554403。结果表明, 多种PCR方法联合使用可以高效的实现对鸭肠炎病毒未知基因的克隆。
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| 关 键 词： | 鸭肠炎病毒 未知基因 简并PCR Targeted gene walking PCR 热不对称交错PCR Long-PCR |
Application of Several PCR Methods in Cloning Unknown Genes of Duck Enteritis Virus |
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| PAN Hua-Qi,CAO Rui-Bing,CHEN Pu-Yan,LIU Lei,WANG Shu-Jin,PAN Guang-Yan and HU Jiang-Chun. Application of Several PCR Methods in Cloning Unknown Genes of Duck Enteritis Virus[J]. Microbiology China, 2009, 36(12): 1842-1848 |
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| Authors: | PAN Hua-Qi CAO Rui-Bing CHEN Pu-Yan LIU Lei WANG Shu-Jin PAN Guang-Yan HU Jiang-Chun |
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| Affiliation: | 1. Institute of Applied Ecology, Chinese Academy Sciences, Shenyang, Liaoning 110016, China; Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture at Nanjing Agricultural University, Nanjing, Jiangsu 210095, China; College of;Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture at Nanjing Agricultural University, Nanjing, Jiangsu 210095, China;Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture at Nanjing Agricultural University, Nanjing, Jiangsu 210095, China;College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu 730070, China;Institute of Applied Ecology, Chinese Academy Sciences, Shenyang, Liaoning 110016, China;College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu 730070, China;Institute of Applied Ecology, Chinese Academy Sciences, Shenyang, Liaoning 110016, China |
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| Abstract: | Degenerate PCR, modified targeted gene walking PCR, modified thermal asymmetric interlaced PCR and Long-PCR were performed with genomic DNA of duck enteritis virus as a template and three fragments (5350 bp, 11083 bp, 2905 bp) of DEV genome were obtained. DNA sequence analysis revealed nine open reading frames encoding UL3, UL4, UL5, UL25, UL26, UL27, UL28, and UL30 gene of DEV, respectively. These sequences have been submitted to GenBank with accession numbers from EF554396 to EF554402. Joint application of several PCR techniques is efficient in cloning unknown genes of duck enteritis virus. |
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| Keywords: | Duck enteritis virus Unknown gene Degenerate PCR Thermal asymmetric interlaced PCR Targeted gene walking PCR Long-PCR |
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