| 鸭瘟病毒北京株UL6和UL7基因的克隆及序列测定 |
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| 引用本文： | 郭霄峰,廖明,辛朝安.鸭瘟病毒北京株UL6和UL7基因的克隆及序列测定[J].畜牧兽医学报,2002,33(6):615-618. |
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| 作者姓名： | 郭霄峰 廖明 辛朝安 |
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| 作者单位： | 华南农业大学动物医学系,广州,510642 |
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| 基金项目： | 广东省自然科学基金项目 ( 994 156 )，华南农业大学校长基金的资助 ( 974 379) |
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| 摘 要： | 依据文献报道的鸭瘟病毒（DPV）的核苷酸序列，设计和合成了二对引物，分别为SP1和SP2，LP1和LP2。北京株鸭瘟病毒于鸭胚中增殖，差速离心和蔗糖密度梯度离心纯化，按酚－氯仿法抽提DNA。然后以DPV DNA为模板进行PCR，分别扩散增出与理论相符的421bp和1209bp的二个DNA片段，并将它们克隆于质粒pGEM-T Easy中。经酶切和质粒PCR，筛选含有目的基因的重组质粒。重组质粒以步移法从双方向测序，获1586bp的核苷酸序列。研究发现这1586bp的核苷酸序列与文献报道的UL6和UL7基因序列的同源性为99％，仅有一个碱基的差异。进一步作氨基酸分析，发现这个碱基所引起的变异为无意义突变（CCT→CCC）。结果表明，鸭瘟病毒的UL6和UL7基因在不同的毒株中高度保守。
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| 关 键 词： | 鸭瘟 病毒 北京株 UL6 UL7 分子克隆 基因测序 |
| 文章编号： | 0366-6964(2002)06-0615-04 |
Cloning and Sequencing of UL6 and UL7 Genes of Duck Plague Virus |
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| GUO Xiao feng,IAO Ming,XIN Chao an.Cloning and Sequencing of UL6 and UL7 Genes of Duck Plague Virus[J].Acta Veterinaria et Zootechnica Sinica,2002,33(6):615-618. |
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| Authors: | GUO Xiao feng IAO Ming XIN Chao an |
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| Abstract: | According to the sequences of Duck Plague Virus(DPV)from GenBank,two pairs of primer,SP1 and SP2,LP1 and LP2,were designed.DPV was cultured in duck embryoes,and the collected allantoic fluid was treateded by differential centrifugation and sucrose density gradient centrifugations.The DNA of DPV was abstracted and was used as the model in PCR.The PCR results revealed that two DNA fragments were about 421bp and 1209bp,which were cloned into plasmid pGEM T Easy.After identification of recombinant by PCR and EcoRI digestion,the inserted DNA fragments were sequenced and sequence alignment showed 99%homology between Beijing strain and American one.The further amino acid analysis demonstrated that the mutation from CCT to CCC was no sense. |
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| Keywords: | Duck Plague Virus Molecular clome Sequencing |
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