Jefferson R A, Kavanagh T A, Bevan M W
Department of Molecular Genetics, Plant Breeding Institute, Trumpington, Cambridge, UK.
EMBO J. 1987 Dec 20;6(13):3901-7. doi: 10.1002/j.1460-2075.1987.tb02730.x.
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.
我们已将大肠杆菌β-葡萄糖醛酸酶基因(GUS)用作基因融合标记,以分析转基因植物中的基因表达。所测试的高等植物缺乏内在的β-葡萄糖醛酸酶活性,因此提高了测量的灵敏度。我们构建了基因融合体,使用花椰菜花叶病毒(CaMV)35S启动子或编码核酮糖二磷酸羧化酶小亚基(rbcS)的基因的启动子来指导转基因植物中β-葡萄糖醛酸酶的表达。使用极少量转基因植物组织的荧光测定法可以准确测量GUS的表达。表达GUS的植物正常、健康且可育。GUS非常稳定,长时间储存后组织提取物仍显示出高水平的GUS活性。组织化学分析已用于证明转基因植物细胞和组织中基因活性的定位。