双顺反子构建体能够轻松检测人细胞中源自酵母人工染色体（YAC）DNA的人囊性纤维化跨膜传导调节因子（CFTR）的表达。
A dicistronic construct allows easy detection of human CFTR expression from YAC DNA in human cells.
作者信息
Vassaux G, Huxley C
机构信息
Department of Biochemistry and Molecular Genetics, Imperial College School of Medicine at St Mary's, Norfolk Place, London W2 1PG, UK.
出版信息
Nucleic Acids Res. 1997 Oct 15;25(20):4167-8. doi: 10.1093/nar/25.20.4167.
Abstract
We have made a dicistronic construct where the picornaviral internal ribosome-entry site (IRES) driving the expression of the beta-geo gene has been inserted into the 3'untranslated region of the human CFTR gene present in a YAC. When introduced into the human cell line Caco-2 expressing the CFTR gene, the expression of the dicistronic gene can be detected by lacZ staining and follows the accumulation of the endogenous CFTR mRNA upon differentiation of the cells. These data demonstrate that this IRES-based approach presents an alternative to mRNA in situ hybridisation and allows detection of expression in an autologous system.
摘要
我们构建了一个双顺反子结构，其中驱动β-geo基因表达的微小核糖核酸病毒内部核糖体进入位点（IRES）已被插入到酵母人工染色体（YAC）中存在的人类CFTR基因的3'非翻译区。当将其导入表达CFTR基因的人类细胞系Caco-2中时，可通过lacZ染色检测双顺反子基因的表达，并且在细胞分化时其表达随内源性CFTR mRNA的积累而变化。这些数据表明，这种基于IRES的方法是mRNA原位杂交的一种替代方法，并且能够在自体系统中检测基因表达。
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