摘要
In the development of techniques by using a bioreactor to produce a special protein,and the establishment of a temporary expression system from primary cell culture oîspecial tissueto analyze the expression regulators and the recombinant gene will greatly expedite the progress ofresearch.This paper is the first report on the establis hment of a temporary expression system fromthe primary culture of chicken oviduet epithelium,which continvously synthesized and secretedovalbumin(OA)during more than two weeks of incubation.When the secretion function of the cellsdecreased,it cauld be restored in most of the cells by adding bormones in the culture medium withinone week of incubation,The OA can be quickly and easily measured by using dot immuofiltrationassay in the cell culture medium because it is a kind of secretary protein.In order to examinewhether the exogenous genes can be expressed in the primary culture cells,the green fluorescentprotein gene(GFPG)was transfected into oviduet cells,and green fluorescence was observed in thecytoplasm of the epithelial cells.These results indicate that the temporary expression system inchicken oviduct epithelium is an effective and convenient system to analyze the expression regulatorsand the recambinant gene.
在开发利用生物反应器生产特定蛋白的研究中,若先用特异组织作原代培养,建立瞬时表达系统,对特定调控元件和融合基因进行分析,可大大缩短研究进程。本文首次报道用鸡输卵管上皮细胞原代培养,建立瞬时表达系统的方法。输卵管细胞体外培养(Fig.1-3),在二,三周时间内仍然保持分泌卵清蛋白的功能,当分泌功能减弱时,若地培液中添加激素,一般一周后大部分细胞又可恢复分泌功能(Fig.4),由于卵清蛋白是一种分泌蛋白,通过斑点免疫渗滤法可迅速简便的从培液中检测到(Fig.4)。为验证培养细胞能否表达外源基因,在原代培养细胞中转染绿色荧光蛋白基因,可在细胞浆中显示绿色荧光(Fig.5)。说明这是一个有效而又方便的检测调控元件的瞬时表达系统。
