GUS fusions: beta‐glucuronidase as a sensitive and versatile gene fusion marker in higher plants.
GUS fusions: beta‐glucuronidase as a sensitive and versatile gene fusion marker in higher plants.
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DOI：
10.1002/j.1460-2075.1987.tb02730.x
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发表时间：
1987-12
期刊：
影响因子：
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通讯作者：
R. Jefferson;T. Kavanagh;M. Bevan
中科院分区：
文献类型：
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作者：
R. Jefferson;T. Kavanagh;M. Bevan
我们使用大肠杆菌β-葡萄糖醛酸酶基因（GUS）作为基因融合标记，用于分析转化植物中的基因表达。测试的高等植物缺乏内在的β-葡萄糖醛酸酶活性，从而提高了测量的灵敏度。我们已经使用花椰菜花叶病毒（CaMV）35 S启动子或来自编码核酮糖二磷酸羧化酶（rbcS）的小亚基的基因的启动子构建了基因融合体，以指导β-葡萄糖醛酸苷酶在转化植物中的表达。GUS的表达可以使用非常少量的转化植物组织的荧光测定法来精确地测量。表达GUS的植物是正常的、健康的和可育的。GUS非常稳定，组织提取物在长期储存后继续显示出高水平的GUS活性。组织化学分析已被用于证明基因活性在转化植物的细胞和组织中的定位。
We have used the Escherichia coli beta‐glucuronidase gene (GUS) as a gene fusion marker for analysis of gene expression in transformed plants. Higher plants tested lack intrinsic beta‐glucuronidase activity, thus enhancing the sensitivity with which measurements can be made. We have constructed gene fusions using the cauliflower mosaic virus (CaMV) 35S promoter or the promoter from a gene encoding the small subunit of ribulose bisphosphate carboxylase (rbcS) to direct the expression of beta‐glucuronidase in transformed plants. Expression of GUS can be measured accurately using fluorometric assays of very small amounts of transformed plant tissue. Plants expressing GUS are normal, healthy and fertile. GUS is very stable, and tissue extracts continue to show high levels of GUS activity after prolonged storage. Histochemical analysis has been used to demonstrate the localization of gene activity in cells and tissues of transformed plants.