綠色熒光蛋白GFP在基因工程中的作用

綠色熒光蛋白（Green fluorescent protein，GFP）是一種在紫外線或藍(lán)光照射下發(fā)出亮綠色熒光的蛋白質(zhì)。最當(dāng)初GFP是從海洋水母維多利亞多管發(fā)光水母中分離出來的。GFP在細(xì)胞和分子生物學(xué)中，常作為報告基因用來檢測轉(zhuǎn)基因生物體中的基因表達(dá)。

綠色熒光蛋白 (GFP) 作為活細(xì)胞和生物體中的通用報告基因具有許多理想的特性。綠色熒光蛋白（GFP）有一個明顯優(yōu)勢是，無需任何底物顯色，在合適波段的激發(fā)光照射下，即可在活細(xì)胞中輕松觀測到GFP表達(dá)。同時對被檢測對象沒有傷害?？梢灾苯佑^測單個細(xì)胞中的基因表達(dá)，無需細(xì)胞裂解和隨后的生化分析，并且可以避免由固定、染色和切片引起的組織變形。發(fā)色團(tuán)的自催化形成和相對的光漂白抗性使GFP成為研究蛋白質(zhì)相互作用、定位和運輸?shù)睦硐霟晒鈽?biāo)簽。GFP的表達(dá)已在大腸桿菌（Escherichia coli）、酵母（yeast）、秀麗隱桿線蟲（Caenorhabditis elegans）、果蠅（Drosophila）、以及各種哺乳動物和植物中得到證實。

GFP在植物中熒光表達(dá)的效果圖

圖一：綠色熒光蛋白GFP在根系的表達(dá)（在析浦GFPfinder-2101RB熒光手電筒照射下）

圖二：綠色熒光蛋白在煙草葉片的表達(dá)（在析浦GFPfinder-2101RB熒光手電筒照射下）

Green fluorescent protein (GFP) is a protein that emits bright green fluorescence under ultraviolet or blue light irradiation. GFP was originally isolated from the marine jellyfish Aequorea Victoria. In cell and molecular biology, GFP is often used as a reporter gene to detect gene expression in genetically modified organisms.

Green fluorescent protein (GFP) has many desirable properties as a universal reporter gene in living cells and organisms. An obvious advantage of green fluorescent protein (GFP) is that it does not require any substrate, and GFP expression can be easily observed in living cells. At the same time, there is no harm to the detected object. Gene expression in single cells can be directly observed without the need for cell lysis and subsequent biochemical analysis, and tissue distortion caused by fixation, staining, and sectioning can be avoided. The autocatalytic formation of the chromophore and relative resistance to photobleaching make GFP an ideal fluorescent label for studying protein interactions, localization, and trafficking. The expression of GFP has been confirmed in Escherichia coli, yeast, Caenorhabditis elegans, Drosophila, and various mammals and plants.