A Preliminary Study on Stable Transfection of EGFP in Buffalo Cumulus Cells

文献类型： 外文期刊

第一作者： Shang, Jiang-Hua

作者： Shang, Jiang-Hua;Yang, Chun-Yan;Qin, Jing;Zheng, Hai-Ying;Huang, Fen-Xiang;Wang, Jian;Liu, Hua-Zhong;Liu, Hua-Zhong

作者机构：

关键词： buffalo;cumulus cells;EGFP;transfection;G418 selection

期刊名称：BUFFALO BULLETIN （ 影响因子：0.172； 五年影响因子：0.234 ）

ISSN： 0125-6726

年卷期： 2013 年 32 卷

页码：

收录情况： SCI

摘要： Transgenic somatic cell nuclear transfer (SCNT) is currently one of effective ways for producing transgenic animals, but the efficiency of the exogenous gene integrating into somatic cells is still low. To obtain stably transfected buffalo somatic cells for transgenic SCNT for further experiments, in this study, buffalo cumulus cells were transfected with pEGFP-N1 by using Lipofectamine 2000 and selected by exposing to G418 (600 mu g/ml) for 3 weeks. Selected transfected cells were imaged under a fluorescence microscope. After 24 hours following transfection, approximate 5% of cells were transiently transfected. Following G418 treatment for 3 weeks, 15% of the survived cells expressed EGFP with fluorescence intensity several folds larger than that in the cells without G418 selection (control). EGFP expressing cells, even part of non-EGFP expressing cells with G418 selection (treatment) were detected to have complex shapes in their morphology which is different from the typical fusiform shape of cumulus cells (control). This study demonstrates that EGFP can successfully be expressed in buffalo cumulus cells and stably transfected cells can be enriched by G418 selection for multiple purposes. Future study should be done to improve the integration rate of exogenous gene and the percentage of stably transfected cells.

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