Amplification and Clone of VP2-4-3 Gene of Very Virulent Infectious Bursal disease virus

SUN Jian—he; JIANG Jing; LU Ping; ZHAO Yu

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August 2002

SUN Jian—he, JIANG Jing, LU Ping and ZHAO Yu. Amplification and Clone of VP2-4-3 Gene of Very Virulent Infectious Bursal disease virus[J]. Virologica Sinica, 2002, 17(4): 358-361.

Citation: SUN Jian—he, JIANG Jing, LU Ping, ZHAO Yu. Amplification and Clone of VP2-4-3 Gene of Very Virulent Infectious Bursal disease virus .VIROLOGICA SINICA, 2002, 17(4) : 358-361.

一步法扩增克隆IBDV上海超强毒VP2-4-3基因

1.
上海交通大学农业与生物学院生物技术研究所，上海201101

摘要

分离、纯化了鸡传染性法氏囊病病毒超强毒(wlBDV)上海株SH95的病毒核酸dsRNA，应用随机引物将 RNA反转录成eDNA，以此为模板一步扩增出A片段前体融合蛋白基因即vP2—4—3基因，将其克隆入pGEM．T载体．并进行序列分析，其与超强毒株HK46的核苷酸序列的同源性达98％，整个基因有5个氨基酸差异，同源性达 99．51％(1007／1012)。
- 鸡传染性法氏囊病病毒超强毒(vvlBDV)
- , 全长eDNA
- , VP2．4．3基因
- , 克隆

Amplification and Clone of VP2-4-3 Gene of Very Virulent Infectious Bursal disease virus

1.
The Institute of Bio Technology ，School of Agriculture，Shanghai JiaoTong University，Shanghai，201101，China

Abstract

The methods of reverse transcription，polymerase chain reaction(PCR)amplification，and cloning of full—length VP2—4—3 gene of a very virulent infectious Bursal disease virus(vvlBDV)strain SH95 were developed．The use of random primer and a reverse transcriptase lacking RNase—H activity produced full—length coding region and non—coding region eDNA copies of the viral genomic segments． The 3060 base—pairs(bp)of VP2—4—3 were amplified by long and accurate PCR in a single step，SUC— cessfully cloned and sequenced revealing their identity of IBDV．
- Very virulent infectious Bursal disease virus(vvlBDV)
- , Full—length eDNA
- , VP2—4—3 gene
- , Clone and sequence analysis

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Amplification and Clone of VP2-4-3 Gene of Very Virulent Infectious Bursal disease virus

1. The Institute of Bio Technology ，School of Agriculture，Shanghai JiaoTong University，Shanghai，201101，China

Abstract: The methods of reverse transcription，polymerase chain reaction(PCR)amplification，and cloning of full—length VP2—4—3 gene of a very virulent infectious Bursal disease virus(vvlBDV)strain SH95 were developed．The use of random primer and a reverse transcriptase lacking RNase—H activity produced full—length coding region and non—coding region eDNA copies of the viral genomic segments． The 3060 base—pairs(bp)of VP2—4—3 were amplified by long and accurate PCR in a single step，SUC— cessfully cloned and sequenced revealing their identity of IBDV．