McInnis R, Perkinson R A, Kuo B A, Norton P A
Roger Williams Medical Center, Providence, RI, USA.
Biochem Mol Biol Int. 1995 Jul;36(3):483-90.
A lacZ cassette was designed to include a synthetic amino terminus optimized for translation in eukaryotic cells, as well as multiple restriction sites for the insertion of heterologous regulatory elements both 5' and 3' of the reporter. The cassette was placed under the control of the metallothionein promoter in combination with the SV40 enhancer and this plasmid was introduced into mammalian cells. High levels of beta-galactosidase were observed in several cell types, demonstrating efficient expression of the reporter. Unexpectedly, most of the chromogenic reaction product appeared to be intra- or peri-nuclear, indicating that the enzyme is similarly localized. The synthetic amino terminus does not resemble known nuclear localization signals and thus may constitute a novel signal.
设计了一个lacZ盒，其中包含一个为在真核细胞中翻译而优化的合成氨基末端，以及多个限制酶切位点，用于在报告基因的5'和3'端插入异源调控元件。该盒置于金属硫蛋白启动子与SV40增强子的控制之下，并将此质粒导入哺乳动物细胞。在几种细胞类型中观察到高水平的β-半乳糖苷酶，表明报告基因有效表达。出乎意料的是，大多数显色反应产物似乎位于核内或核周，这表明该酶也定位于此。合成氨基末端与已知的核定位信号不同，因此可能构成一种新的信号。
