鸭肝炎病毒A66株部分cDNA文库的构建与序列分析

›› 2007, Vol. 34 ›› Issue (9): 84-86.

鸭肝炎病毒A₆₆株部分cDNA文库的构建与序列分析

赵瑞宏1，张小飞2,3，温立斌2，潘孝成1

1.安徽省农科院畜牧兽医研究所，合肥 230031；2.江苏省农科院兽医研究所，南京 210014；3.南京天邦生物科技有限公司，南京 211102

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2007-09-20 发布日期:2007-09-20

Construction and Analysis of Partial cDNA Library of Duck Hepatitis Virus Strain A₆₆

ZHAO Ruihong1,ZHANG Xiaofei2,3,WEN Libin2,PAN Xiaocheng1

1.Institute of Animal Husbandry and Veterinary Medicine, Anhui Academy of Agricultural Sciences, Hefei 230031,China;2.Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;3.Nanjing Techbank Bio-industry Co Ltd, Nanjing 211102, China

Received:1900-01-01 Revised:1900-01-01 Online:2007-09-20 Published:2007-09-20

摘要/Abstract

摘要： 本研究根据已发表的小核糖核酸病毒（picornavirus）核苷酸序列设计4对引物。以RNAiso Reagent试剂抽提含DHV A₆₆鸡胚尿囊液总RNA ，用TakaRa RNA PCR Kit（AMV）Ver3.0试剂盒进行RT-PCR扩增，将所得PCR产物回收并克隆于pMD18-T 载体、转化感受态细菌DH5а，利用氨苄青霉素筛选阳性克隆子, 经EcoRⅠ和SalⅠ双酶切鉴定重组质粒。取重组阳性质粒进行测序，测序结果与近期GenBank登录的3株Ⅰ型DHV核苷酸序列比对，结果显示与03D株、H株、5886株鸭肝炎病毒的核酸序列同源性分别为96.8%、96.3%、96.2%，表明成功构建了DHV A₆₆株部分cDNA文库。

关键词: 鸭肝炎病毒; A₆₆株; cDNA文库; 序列分析

Abstract: Based on the published sequences of picornavirus, 4 pairs of RT-PCR primer were designed. Total RNA were extracted from chick embryo allantoic fluid by RNAiso Reagent,one cDNA fragment were successfully amplified from total RNA extracted from DHV A₆₆chick embryo allantoic fluid by using TakaRa
RNA PCR Kit（AMV）Ver3.0. After ligating cDNA to the plasmid vector pMD18T and introducing into E. coli DH5а, the recombinant plasmids were screened by ampicillin and checked by the restriction enzyme EcoRⅠ and SalⅠ digestion. Then the recombinant plasmids were sequenced. Compare and analyse of homologies of nucleotide showed that identities of DHV A₆₆and 3 published DHV were up to 96.2%～96.8%. This indicated partial cDNA library of DHV A₆₆ strain were constructed successfully.

Key words: duck hepatitis virus(DHV); A₆₆strain; cDNA library; sequence analysis

中图分类号:

S852.65+9.6

赵瑞宏;张小飞;温立斌;潘孝成. 鸭肝炎病毒A₆₆株部分cDNA文库的构建与序列分析[J]. , 2007, 34(9): 84-86.

ZHAO Ruihong;ZHANG Xiaofei;WEN Libin;PAN Xiaocheng. Construction and Analysis of Partial cDNA Library of Duck Hepatitis Virus Strain A₆₆[J]. , 2007, 34(9): 84-86.

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鸭肝炎病毒A₆₆株部分cDNA文库的构建与序列分析

Construction and Analysis of Partial cDNA Library of Duck Hepatitis Virus Strain A₆₆

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