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盘基网柄菌的一种反式作用调节突变体在真核表达系统中的应用。

Use of a transactive regulatory mutant of Dictyostelium discoideum in a eucaryotic expression system.

作者信息

Wetterauer B W, Salger K, MacWilliams H K

机构信息

Zoologisches Institut, Ludwig-Maximilians Universität München, Germany.

出版信息

Nucleic Acids Res. 1993 Mar 25;21(6):1397-401. doi: 10.1093/nar/21.6.1397.

Abstract

The discoidin proteins of Dictyostelium discoideum are highly expressed during development. The Disc I gamma promoter allows the regulation of heterologous protein expression by experimental conditions. We report conditions under which the promoter activity is efficiently repressed during growth in the wildtype strain AX2. In addition we show that a mutant which overexpresses the discoidins also overexpresses the reporter genes beta-galactosidase, luciferase and CAT 10- to 100-fold when these are placed under the control of a Disc I gamma promoter. This system may be generally useful for the overexpression of genes in Dictyostelium, both for functional studies in vivo and for the production of heterologous proteins for purification.

摘要

盘基网柄菌的盘状球蛋白在发育过程中高度表达。盘状球蛋白Iγ启动子可通过实验条件调控异源蛋白表达。我们报道了在野生型菌株AX2生长期间有效抑制启动子活性的条件。此外,我们还表明,当报告基因β-半乳糖苷酶、荧光素酶和氯霉素乙酰转移酶置于盘状球蛋白Iγ启动子控制下时,过表达盘状球蛋白的突变体也会使其过表达10至100倍。该系统可能普遍适用于盘基网柄菌中基因的过表达,既用于体内功能研究,也用于生产用于纯化的异源蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/faf9/309324/67315fb9f28d/nar00055-0071-a.jpg

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