家蚕细胞中功能性S-II的产生。

Production of functional S-II in Bombyx mori cells.

作者信息

Hirashima S, Horikoshi N, Sekimizu K, Natori S

机构信息

Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.

出版信息

Biochem Biophys Res Commun. 1989 May 15;160(3):1093-9. doi: 10.1016/s0006-291x(89)80115-9.

DOI:10.1016/s0006-291x(89)80115-9

PMID:2730640

Abstract

A cDNA for murine transcription factor S-II (Hirashima et al., J. Biol. Chem (1988) 263, 3858-3863) was inserted into a silkworm baculovirus vector and expressed in Bm-N cells, a cell line of Bombyx mori maintained in the laboratory. Recombinant S-II was purified from virus-infected cell extracts to near homogeneity by ammonium sulfate fractionation, and chromatographies on DEAE-cellulose and phosphocellulose. About 60 micrograms of recombinant S-II was obtained from 1 g of virus-infected cells. The molecular mass and specific activity of recombinant S-II were exactly the same as those of authentic S-II purified from Ehrlich ascites tumor cells.

摘要

将小鼠转录因子S-II的cDNA（平岛等人，《生物化学杂志》（1988年）263卷，3858 - 3863页）插入家蚕杆状病毒载体，并在实验室保存的家蚕细胞系Bm - N细胞中表达。通过硫酸铵分级沉淀以及DEAE - 纤维素和磷酸纤维素柱层析，从病毒感染的细胞提取物中纯化重组S-II至接近均一状态。从1克病毒感染细胞中获得了约60微克的重组S-II。重组S-II的分子量和比活性与从艾氏腹水瘤细胞中纯化得到的天然S-II完全相同。