稳定番茄黄叶卷曲 中国病毒 传染性克隆 通过微同类学介导的末端连接
Anxiang Wang1, Tian Meng1, Ju Wang1
1State Key Laboratory of Biocatalysis and Enzyme Engineering, College of Life Sciences, Hubei University, Wuhan, China.
Plant biotechnology journal
|November 30, 2025
概括
一种新的微同质介导端结合 (MMEJ) 系统稳定了细菌中的西红黄色叶子卷曲中国病毒 (TYLCCNV) 传染性克隆. 这种方法确保了植物中有效的病毒基因组释放,改善了DNA病毒克隆策略.
科学领域:
- 植物病毒学 植物病毒学
- 分子生物学分子生物学
- 农业科学 农业科学
背景情况:
- 番茄黄叶卷曲中国病毒 (TYLCCNV) 是一个重要的农业病原体,也是圆形单链DNA (cssDNA) 病毒的模型.
- 目前用于构建TYLCCNV传染性克隆的方法涉及Agrobacterium中的并联病毒基因组,这往往导致在培养过程中病毒性丧失.
研究的目的:
- 为TYLCCNV和其他植物DNA病毒开发稳定的传染性克隆系统.
- 为了克服cssDNA病毒传染性克隆在原生宿主中的不稳定性.
主要方法:
- 设计了一种微同质介导端结合 (MMEJ) 系统,以破坏病毒复制蛋白 (Rep) 编码序列.
- 在Rep基因中插入了一个I-SceI识别位点,旁边是微同类区域.
- 利用植物中的短暂I-SceI表达来触发MMEJ修复,切除I-SceI部位并释放功能性病毒基因组.
主要成果:
- 基于MMEJ的系统成功地稳定了TYLCCNV传染性克隆在Agrobacterium.
- 植物中I-SceI的短暂表达导致了工程序列的无去除.
- 在植物中有效释放了功能性TYLCCNV基因组,恢复了毒性.
结论:
- 开发的MMEJ系统为稳定植物DNA病毒的传染性克隆提供了强大的方法.
- 这种方法为构建和操纵cssDNA病毒基因组提供了一个广泛适用的策略.
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