Cloning and Sequence Analysis of N gene encoding Akabane virus  Nucleotide Capsid Protein

HUA Qun-yi; ZHANG Nian-zu; DONG Jun; YANG Jing-yan; XU Zi-zhong; YANG Yun-qin

PDF
Cite
Share

facebook

twitter

google

linkedin

February 2004

HUA Qun-yi, ZHANG Nian-zu, DONG Jun, YANG Jing-yan, XU Zi-zhong and YANG Yun-qin. Cloning and Sequence Analysis of N gene encoding Akabane virus Nucleotide Capsid Protein[J]. Virologica Sinica, 2004, 19(1): 18-21.

Citation: HUA Qun-yi, ZHANG Nian-zu, DONG Jun, YANG Jing-yan, XU Zi-zhong, YANG Yun-qin. Cloning and Sequence Analysis of N gene encoding Akabane virus Nucleotide Capsid Protein .VIROLOGICA SINICA, 2004, 19(1) : 18-21.

赤羽病病毒核蛋白基因克隆和序列分析*

花群义 ^1*,, ,
张念祖 ² ,
杨晶焰 ¹ ,
徐自忠 ¹ ,
杨云庆 ¹ ,

1.
1. 云南出入境检验检疫局技术中心云南昆明 650228
2.
农业部热带亚热带动物病毒学重点开放实验室, 云南昆明 650224

通讯作者： 花群义, ;

摘要

参考GeneBank发表的赤羽病病毒（Akabane virus,AKAV）的核蛋白基因（SmRNA）序列，设计合成一对引物，从分离自牛体的AKAV BHK21细胞培养物中提取总RNA，对AKAV核蛋白基因进行RT-PCR扩增，产物经琼脂糖电泳分析，呈现一条约696bp的条带，回收纯化后，将其克隆至pMD18-T质粒载体中，然后进行核苷酸序列分析。与GenBank中报道的多株AKAV编码核衣壳蛋白（N）的SmRNA基因比较后发现，与其它株的核苷酸的同源性为94.2%~98.3%，推导的氨基酸的同源性为97.6%~100%，证实为AKV的N基因。为生产AKAV特异性核蛋白抗原、免疫血清学诊断试剂的制备和分子生物学研究打下了坚实基础。
- 赤羽病病毒
- , 核蛋白基因
- , 克隆与序列分析

Cloning and Sequence Analysis of N gene encoding Akabane virus Nucleotide Capsid Protein

1.
1. Technology Center of Yunnan Entry-Exit Inspection and Quarantine, Kunming 650228, China
2.
The key Opening Laboratory of Tropical and Subtropical Animal Virology of Ministry of Agriculture, Kunming 650224, China

Corresponding author: HUA Qun-yi,

Abstract

The nucleocapsid protein (N) gene of Akabane virus strain YN isolated from bovine was amplified by RT-PCR. The product of 696 bp fragment was obtained as expected. The amplified fragment was then cloned into the pDM18-T vector and confirmed by PCR, endonuclease analysis, and sequencing. The results demonstrated that nucleotide homology with other published AKAV N genes was between 94.2%~98.3%, and the deduced amino acid homology was between 97.6%~99.5%, respectively. Therefore, nucleotide capsid protein gene of AKAV was conserved. Phylogenetic tree constructed with 17 reference strains of 696 bp of the N gene showed that AKAV-YN strain is related to Tinaroo strain.
- Akabane virus
- , N gene Cloning
- , Sequence analysis

References
Proportional views

PDF

Article Metrics

Article views(4059) PDF downloads(908) Cited by(0)

Proportional views

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

Cloning and Sequence Analysis of N gene encoding Akabane virus Nucleotide Capsid Protein

Corresponding author: HUA Qun-yi,

1. 1. Technology Center of Yunnan Entry-Exit Inspection and Quarantine, Kunming 650228, China
2. The key Opening Laboratory of Tropical and Subtropical Animal Virology of Ministry of Agriculture, Kunming 650224, China

Abstract: The nucleocapsid protein (N) gene of Akabane virus strain YN isolated from bovine was amplified by RT-PCR. The product of 696 bp fragment was obtained as expected. The amplified fragment was then cloned into the pDM18-T vector and confirmed by PCR, endonuclease analysis, and sequencing. The results demonstrated that nucleotide homology with other published AKAV N genes was between 94.2%~98.3%, and the deduced amino acid homology was between 97.6%~99.5%, respectively. Therefore, nucleotide capsid protein gene of AKAV was conserved. Phylogenetic tree constructed with 17 reference strains of 696 bp of the N gene showed that AKAV-YN strain is related to Tinaroo strain.