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水稻(Oryza sativa L.)中雌激素诱导的 GFP 表达模式。

Estrogen-inducible GFP expression patterns in rice (Oryza sativa L.).

机构信息

Division of Plant Sciences, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan.

出版信息

Plant Cell Rep. 2011 Apr;30(4):529-38. doi: 10.1007/s00299-010-0963-0. Epub 2010 Dec 8.

Abstract

We investigated estrogen-inducible green fluorescent protein (GFP) expression patterns using an estrogen receptor fused chimeric transcription activator, XVE, in the monocotyledonous model plant rice (Oryza sativa L.). This system has been shown to be an effective chemical-inducible gene expression system in Arabidopsis and has been applied to other plants in order to investigate gene functions or produce marker-free transgenic plants. However, limited information is available on the correlation between inducer concentration and the expression level of the gene induced in monocots. Here, we produced a transgenic rice integrated estrogen-inducible GFP expression vector, pLex:GFP, and investigated dose-response and time-course patterns of GFP induction in rice calli and seedlings for the first time. With 17-β-estradiol treatment at >5 μM, GFP signals were detected in the entire surface of calli within 2 days of culture. Highest GFP signals were extended for 8 days with estradiol treatment at 25 μM. In three-leaf-stage seedlings, GFP signals in the leaves of pLex:GFP-integrated transgenic lines were weaker than those in the leaves of p35S:GFP-integrated transgenic lines. However, GFP signals in the roots of pLex:GFP- and p35S:GFP-integrated transgenic lines were similar with estradiol treatment at >10 μM. With regard to controlling appropriate gene expression, these results might provide helpful indications on estradiol treatment conditions to be used for the XVE system in rice and other monocots.

摘要

我们使用雌激素受体融合嵌合转录激活子 XVE 研究了单子叶模式植物水稻(Oryza sativa L.)中雌激素诱导的绿色荧光蛋白(GFP)表达模式。该系统已被证明是拟南芥中一种有效的化学诱导基因表达系统,并已应用于其他植物,以研究基因功能或产生无标记转基因植物。然而,关于诱导剂浓度与单子叶植物中诱导基因表达水平之间的相关性的信息有限。在这里,我们首次产生了一种整合雌激素诱导 GFP 表达载体 pLex:GFP 的转基因水稻,并研究了水稻愈伤组织和幼苗中 GFP 诱导的剂量反应和时间进程模式。用 17-β-雌二醇处理>5 μM,在培养的第 2 天,GFP 信号在愈伤组织的整个表面都能检测到。用 25 μM 的雌二醇处理,最高 GFP 信号可延长 8 天。在三叶期幼苗中,pLex:GFP 整合转基因系叶片中的 GFP 信号比 p35S:GFP 整合转基因系叶片中的弱。然而,用>10 μM 的雌二醇处理后,pLex:GFP 和 p35S:GFP 整合的转基因系的根中的 GFP 信号相似。关于控制适当的基因表达,这些结果可能为 XVE 系统在水稻和其他单子叶植物中使用雌二醇处理条件提供有用的指示。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8060/3056999/6b4156067a86/299_2010_963_Fig1_HTML.jpg

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