Development of a GFP expression vector for Cucurbit chlorotic yellows virus.

Development of a GFP expression vector for Cucurbit chlorotic yellows virus.
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葫芦褪绿黄化病毒GFP表达载体的研制

DOI:
10.1186/s12985-018-1004-9
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发表时间:
2018-05-24
期刊:
影响因子:
4.8
通讯作者:
Shi Y
Shi Y
中科院分区:
医学3区
文献类型:
--
作者:
Wei Y;Han X;Wang Z;Gu Q;Li H;Chen L;Sun B;Shi Y

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背景:瓜类褪绿黄化病毒(Cucurbit chlorotic yellowsvirus,CCYV)是一种二分蔓状病毒,可引起瓜类叶片褪绿叶斑和黄化症状。我们以前开发了CCYV的感染性克隆。研究结果:我们利用CCYV RNA 2 cDNA双切的“添加基因”策略构建了CCYV绿色荧光蛋白(GFP)表达载体。用不同的启动子构建了三个得到的克隆,pCCYVGFPSGC、pCCYVGFPCGC和pCCYVGFPCGS,所述启动子用于启动GFP和CP表达。在25 dpi时,GFP荧光不仅在叶脉中而且在周围细胞中可检测到。pCCYVGFPCGC感染黄瓜叶片后25 dpi表现为细胞铺展,而pCCYVGFPSGC和pCCYVGFPCGS主要以单细胞形式存在。进一步观察pCCYVGFPCGCGFP在30 dpi,40 dpi,和50 dpi的表达表明韧皮部限制定位在系统leafs.Conclusions:我们开发了CCYV GFP表达载体,将是有用的,为进一步研究CCYV在葫芦运动。
Background:Cucurbit chlorotic yellows virus (CCYV), a bipartite crinivirus, causes chlorotic leaf spots and yellowing symptoms on cucurbit leaves. We previously developed an infectious clone of CCYV. Limited work has been conducted on the construction of a crinivirus green fluorescence protein (GFP) expression vector to date.Finding:We constructed a CCYV GFP expression vector using the "add a gene" strategy based on CCYV RNA2 cDNA constrcut. Three resultant clones, pCCYVGFPSGC, pCCYVGFPCGC, and pCCYVGFPCGS,were constructed with different promoters used to initiate GFP and CP expression. At 25 dpi GFP fluorescence was detectable not only in leaf veins but also in the surrounding cells. pCCYVGFPCGC-infected cucumber leaves exhibited cell spread at 25 dpi, whereas pCCYVGFPSGCand pCCYVGFPCGSwere mainly found in single cells. Further observation of pCCYVGFPCGCGFP expression at 30 dpi, 40 dpi, and 50 dpi showed phloem-limited localization in the systemic leaves.Conclusions:We developed of a CCYV GFP expression vector that will be useful for further study of CCYV movement in cucurbits.
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发表时间: 2004-04-25
期刊: VIROLOGY
影响因子: 3.7
作者:
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