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一种用于优化酵母-哺乳动物细胞融合条件的简易测定法。

A simple assay for optimizing yeast-mammalian cell fusion conditions.

作者信息

Markie D

机构信息

Paediatric Research Unit, UMDS, Guy's Hospital, London.

出版信息

Mol Biotechnol. 1996 Oct;6(2):99-104. doi: 10.1007/BF02740766.

Abstract

Polyethylene glycol (PEG)-induced cell fusion can be a useful method for the transfer of yeast artificial chromosomes (YACs) from yeast spheroplasts to mammalian cells in culture, although success varies between recipient cell types. Experiments aimed at determining optimum fusion conditions can also be very time-consuming. To minimize this difficulty, a reporter plasmid has been constructed that allows yeast-mammalian cell fusion rates to be determined within 3 d. The speed and sensitivity of the assay should allow a more systematic evaluation of cell lines for their capacity to fuse with yeast, and for rapid optimization of fusion parameters.

摘要

聚乙二醇(PEG)诱导的细胞融合可能是一种将酵母人工染色体(YACs)从酵母原生质体转移到培养的哺乳动物细胞中的有用方法,尽管不同受体细胞类型的成功率有所不同。旨在确定最佳融合条件的实验也可能非常耗时。为了尽量减少这一困难,构建了一种报告质粒,可在3天内确定酵母-哺乳动物细胞融合率。该检测方法的速度和灵敏度应能更系统地评估细胞系与酵母融合的能力,并能快速优化融合参数。

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