Sequence Analysis of the HC-pro gene of Potato Virus Y N strain and its Expression in E .coli

WANG Rui; WU Yun-feng*; HAO Xing-an; FU Hong-qi; YANG Dong; FAN Bing

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June 2005

WANG Rui, WU Yun-feng*, HAO Xing-an, FU Hong-qi, YANG Dong and FAN Bing. Sequence Analysis of the HC-pro gene of Potato Virus Y N strain and its Expression in E .coli[J]. Virologica Sinica, 2005, 20(3): 320-322.

Citation: WANG Rui, WU Yun-feng*, HAO Xing-an, FU Hong-qi, YANG Dong, FAN Bing. Sequence Analysis of the HC-pro gene of Potato Virus Y N strain and its Expression in E .coli .VIROLOGICA SINICA, 2005, 20(3) : 320-322.

马铃薯Y病毒N株HC-pro基因的序列分析与表达

1.
西北农林科技大学植保学院陕西省农业分子生物重点实验室陕西杨凌

摘要

马铃薯Y病毒(PotatovirusY,PVY)是马铃薯Y病毒属(Potyvirus)的典型成员,是烟草、马铃薯和辣椒等茄科作物病毒病的主要病原之一,分布于世界各地。近年烟草上的PVY在全国范围内特别是陕西、黄淮和东北烟区流行呈上升趋势,且以PVY坏死株系(PVY N)为主,重病田发生率达40%以上,并常与...

Sequence Analysis of the HC-pro gene of Potato Virus Y N strain and its Expression in E .coli

1.
College of Plant Protect ion Nort hwest A &F University , Shaanxi Key L aboratory for Agricultural Molecular Biotechnology ,Shaanx i , Yangling 712100 , China

Abstract

The HC-pro gene was amplified by RT-PCR from total RNA of tobacco leaves infected with a N strain of Potato virus Y in Shaanxi, and cloned into the PMD 18-T vector. This HC-pro gene is consisted of 1371 nucleotides, encoding 457 amino acids. It shared the sequence homology of 82.5%~96.4 % nucleotide acid and 92.5%~98.0% in amino acids compared to 9 species of PVY N HC-pro abroad.The HC-pro gene was inserted into prokaryotic expressing vector pBV221, to obtain pBVHC recombinant plasmid in E.coli BL21. SDS-PAGE indicated that HC-pro proteins are successfully expressed in E.coli, Western blotting analysis demonstrated that the antibody against the expressed HC-pro can be used to identify the infected plants .

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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

Sequence Analysis of the HC-pro gene of Potato Virus Y N strain and its Expression in E .coli

1. College of Plant Protect ion Nort hwest A &F University , Shaanxi Key L aboratory for Agricultural Molecular Biotechnology ,Shaanx i , Yangling 712100 , China

Abstract: The HC-pro gene was amplified by RT-PCR from total RNA of tobacco leaves infected with a N strain of Potato virus Y in Shaanxi, and cloned into the PMD 18-T vector. This HC-pro gene is consisted of 1371 nucleotides, encoding 457 amino acids. It shared the sequence homology of 82.5%~96.4 % nucleotide acid and 92.5%~98.0% in amino acids compared to 9 species of PVY N HC-pro abroad.The HC-pro gene was inserted into prokaryotic expressing vector pBV221, to obtain pBVHC recombinant plasmid in E.coli BL21. SDS-PAGE indicated that HC-pro proteins are successfully expressed in E.coli, Western blotting analysis demonstrated that the antibody against the expressed HC-pro can be used to identify the infected plants .