通过DEAE-葡聚糖介导的DNA摄取与渗透休克程序相结合对小鼠淋巴细胞进行DNA转染。

DNA transfection of mouse lymphoid cells by the combination of DEAE-dextran-mediated DNA uptake and osmotic shock procedure.

作者信息

Takai T, Ohmori H

机构信息

Department of Biotechnology, Faculty of Engineering, Okayama University, Japan.

出版信息

Biochim Biophys Acta. 1990 Jan 30;1048(1):105-9. doi: 10.1016/0167-4781(90)90029-2.

DOI:10.1016/0167-4781(90)90029-2

PMID:2297528

Abstract

Several mouse lymphoid cell lines were efficiently transfected with plasmid DNA by a novel method combining DEAE-dextran-mediated DNA uptake and osmotic shock procedure. The cells were first incubated with DNA-DEAE-dextran complex, treated with hypertonic Tris-HCl buffer containing 0.5 M sucrose and 10% poly(ethylene glycol), and then exposed to hypotonic RPMI 1640 medium. This transfection protocol exhibited maximal frequencies of 0.3% and 3.10(-5) for transient gene expression and stable transformation in P3-NSI/1-Ag4-1 cells, respectively.

摘要

通过一种将DEAE-葡聚糖介导的DNA摄取与渗透压休克程序相结合的新方法，几种小鼠淋巴细胞系被高效地用质粒DNA转染。细胞首先与DNA-DEAE-葡聚糖复合物孵育，用含有0.5M蔗糖和10%聚乙二醇的高渗Tris-HCl缓冲液处理，然后暴露于低渗RPMI 1640培养基中。对于P3-NSI/1-Ag4-1细胞中的瞬时基因表达和稳定转化，该转染方案分别显示出最大频率为0.3%和3.1×10⁻⁵ 。