本文選題：mCherry + 紅色熒光蛋白　； 參考：《植物研究》2017年02期

【摘要】：熒光蛋白在生物學(xué)研究中具有廣泛的應(yīng)用和重要的作用,其中紅色熒光蛋白mCherry因其顏色和良好的特性,對于植物基因研究具有重要的使用價值,本研究將mCherry基因構(gòu)建到pBI121植物表達(dá)載體系統(tǒng)中,構(gòu)建了pBI121MCS-mCherry載體。利用基因槍轉(zhuǎn)化法轉(zhuǎn)入洋蔥表皮進行表達(dá)驗證,顯微鏡觀察結(jié)果顯示整個洋蔥細(xì)胞具有紅色熒光,證明該載體能夠在植物細(xì)胞中表達(dá)紅色熒光蛋白。利用雙酶切連接法將轉(zhuǎn)錄因子BpMYB4基因構(gòu)建到該載體上,得到融合表達(dá)載體pBI121MCS-mCherry-BpMYB4,在洋蔥表皮中表達(dá),結(jié)果顯示細(xì)胞核具有紅色熒光,證明該載體能夠準(zhǔn)確表達(dá)融合蛋白,進行亞細(xì)胞定位。同時融合基因時不再需要中間載體,構(gòu)建簡便,引入的KpnⅠ酶切位點,增加了可選擇性。因此該載體可用于植物基因表達(dá)定位及轉(zhuǎn)基因植株篩選研究中,為今后的白樺基因組學(xué)研究提供了材料。
[Abstract]:Fluorescent protein (mCherry) has been widely used and played an important role in biological research. Red fluorescent protein (mCherry) has important application value for plant gene research because of its color and good characteristics. In this study, mCherry gene was constructed into pBI121 plant expression vector system and pBI121MCS-mCherry vector was constructed. The results of microscopical observation showed that the whole onion cells had red fluorescence, which proved that the vector could express red fluorescent protein in plant cells. The transcription factor BpMYB4 gene was constructed into the vector by double enzyme digestion, and the fusion expression vector pBI121MCS-mCherry-BpMYB4 was obtained, which was expressed in onion epidermis. The results showed that the cell nucleus had red fluorescence, which proved that the vector could express the fusion protein accurately. Subcellular localization was performed. At the same time, the intermediate vector is no longer needed in the fusion gene, so it is easy to construct, and the Kpn 鈪，

本文編號：1896984