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Genetic transformation of HeLa cells by Agrobacterium.农杆菌介导的HeLa细胞基因转化
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植物基因对根癌土壤杆菌的表达反应。

Plant gene expression response to Agrobacterium tumefaciens.

作者信息

Ditt R F, Nester E W, Comai L

机构信息

Department of Botany, University of Washington, Seattle, WA 98195-5325, USA.

出版信息

Proc Natl Acad Sci U S A. 2001 Sep 11;98(19):10954-9. doi: 10.1073/pnas.191383498. Epub 2001 Sep 4.

DOI:10.1073/pnas.191383498
PMID:11535836
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC58580/
Abstract

To elucidate the nature of plant response to infection and transformation by Agrobacterium tumefaciens, we compared the cDNA-amplified fragment length polymorphism (AFLP) pattern of Agrobacterium- and mock-inoculated Ageratum conyzoides plant cell cultures. From 16,000 cDNA fragments analyzed, 251 (1.6%) were differentially regulated (0.5% down-regulated) 48 h after cocultivation with Agrobacterium. From 75 strongly regulated fragments, 56 were already regulated 24 h after cocultivation. Sequence similarities were obtained for 20 of these fragments, and reverse transcription-PCR analysis was carried out with seven to confirm their cDNA-AFLP differential pattern. Their sequence similarities suggest a role for these genes in signal perception, transduction, and plant defense. Reverse transcription-PCR analysis indicated that four genes involved in defense response are regulated in a similar manner by nonpathogenic bacteria, whereas one gene putatively involved in signal transduction appeared to respond more strongly to Agrobacterium. A nodulin-like gene was regulated only by Agrobacterium. These results demonstrate a rapid plant cell response to Agrobacterium infection, which overlaps a general response to bacteria but also has Agrobacterium-specific features.

摘要

为阐明植物对根癌农杆菌感染和转化的反应本质,我们比较了经农杆菌接种和模拟接种的藿香蓟植物细胞培养物的cDNA扩增片段长度多态性(AFLP)图谱。在分析的16000个cDNA片段中,与农杆菌共培养48小时后,有251个(1.6%)呈现差异表达(0.5%下调)。在75个强差异表达片段中,有56个在共培养24小时后就已出现差异表达。其中20个片段获得了序列相似性,并对7个片段进行了逆转录-聚合酶链反应(RT-PCR)分析以确认其cDNA-AFLP差异模式。它们的序列相似性表明这些基因在信号感知、转导和植物防御中发挥作用。RT-PCR分析表明,4个参与防御反应的基因受非致病细菌的调控方式相似,而1个推测参与信号转导的基因对农杆菌的反应似乎更强。一个类结瘤素基因仅受农杆菌调控。这些结果证明了植物细胞对农杆菌感染的快速反应,这种反应与对细菌的一般反应重叠,但也具有农杆菌特异性特征。