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从马铃薯X病毒韩国株系和轻度斑驳株系产生感染性转录本。

Generation of infectious transcripts from Korean strain and mild mottle strain of potato virus X.

作者信息

Choi Sun Hee, Ryu Ki Hyun

机构信息

Plant Virus GenBank, Division of Environmental and Life Sciences, Seoul Women's University, Seoul, 139-774, Republic of Korea.

出版信息

J Microbiol. 2008 Oct;46(5):502-7. doi: 10.1007/s12275-008-0078-2. Epub 2008 Oct 31.

Abstract

Full-length cDNAs of two different strains of Potato virus X (PVX-Kr and PVX-Mo) have been directly amplified by long template reverse transcription polymerase chain reaction (RT-PCR) using the 5'-end primer containing a SP6 or T7 RNA promoter sequence and the virus-specific 3'-end primer, and then constructed in plasmid vectors. Capped in vitro transcripts from cloned full-length cDNAs as well as those RT-PCR amplicons proved to be infectious systemically on tobacco plants. Symptom expression on tobacco plants from PVX-Mo transcripts was faster and severer than that from PVX-Kr. In replication stability test of transcripts derived from PVX clones, progeny viruses showed stable replication according to sequencing through passages. This highly infectious transcript system from the full-length cDNA clones for PVX can be useful for recombinant molecules for functional analysis of viral proteins in plant-virus interaction study as well as for expression of foreign protein in planta.

摘要

利用含有SP6或T7 RNA启动子序列的5'端引物和病毒特异性3'端引物,通过长模板逆转录聚合酶链反应(RT-PCR)直接扩增了两种不同马铃薯X病毒株(PVX-Kr和PVX-Mo)的全长cDNA,然后构建到质粒载体中。克隆的全长cDNA以及那些RT-PCR扩增产物的加帽体外转录本在烟草植株上被证明具有系统感染性。PVX-Mo转录本在烟草植株上引起的症状表达比PVX-Kr的更快、更严重。在PVX克隆转录本的复制稳定性测试中,子代病毒通过传代测序显示出稳定的复制。这种来自PVX全长cDNA克隆的高感染性转录本系统可用于植物-病毒相互作用研究中病毒蛋白功能分析的重组分子,以及用于在植物中表达外源蛋白。

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