Andersen J B, Sternberg C, Poulsen L K, Bjorn S P, Givskov M, Molin S
Department of Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark.
Appl Environ Microbiol. 1998 Jun;64(6):2240-6. doi: 10.1128/AEM.64.6.2240-2246.1998.
Use of the green fluorescent protein (Gfp) from the jellyfish Aequorea victoria is a powerful method for nondestructive in situ monitoring, since expression of green fluorescence does not require any substrate addition. To expand the use of Gfp as a reporter protein, new variants have been constructed by the addition of short peptide sequences to the C-terminal end of intact Gfp. This rendered the Gfp susceptible to the action of indigenous housekeeping proteases, resulting in protein variants with half-lives ranging from 40 min to a few hours when synthesized in Escherichia coli and Pseudomonas putida. The new Gfp variants should be useful for in situ studies of temporal gene expression.
利用来自维多利亚多管水母的绿色荧光蛋白（Gfp）是一种用于非破坏性原位监测的强大方法，因为绿色荧光的表达不需要添加任何底物。为了扩大Gfp作为报告蛋白的用途，通过在完整Gfp的C末端添加短肽序列构建了新的变体。这使得Gfp易受内源管家蛋白酶的作用，导致在大肠杆菌和恶臭假单胞菌中合成时，蛋白质变体的半衰期从40分钟到几小时不等。新的Gfp变体应该可用于基因表达时间的原位研究。
