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在利用瞬时基因表达的调控研究中,将人生长激素作为报告基因。

Human growth hormone as a reporter gene in regulation studies employing transient gene expression.

作者信息

Selden R F, Howie K B, Rowe M E, Goodman H M, Moore D D

出版信息

Mol Cell Biol. 1986 Sep;6(9):3173-9. doi: 10.1128/mcb.6.9.3173-3179.1986.

Abstract

The human growth hormone (hGH) transient assay system described here is based on the expression of hGH directed by cells transfected with hGH fusion genes. Levels of secreted hGH in the medium, measured by a simple radioimmunoassay, are proportional to both levels of cytoplasmic hGH mRNA and the amount of transfected DNA. The system is extremely sensitive, easy to perform, and is qualitatively different from other transient expression systems in that the medium is assayed and the cells themselves are not destroyed. The hGH transient assay system is appropriate for analyses of regulation of gene expression and was utilized here to investigate the effect of the simian virus 40 enhancer on the herpes simplex virus thymidine kinase promoter and the effect of zinc on the mouse metallothionein-I promoter. The expression of hGH can also be used as an internal control to monitor transfection efficiency along with any other transient expression system. All cell types tested thus far (including AtT-20, CV-1, GC, GH4, JEG, L, and primary pituitary cells) were able to secrete hGH into the medium.

摘要

本文所述的人生长激素(hGH)瞬时检测系统基于用hGH融合基因转染的细胞所指导的hGH表达。通过简单的放射免疫测定法测量培养基中分泌的hGH水平,其与细胞质hGH mRNA水平和转染DNA的量均成正比。该系统极其灵敏,易于操作,并且与其他瞬时表达系统在性质上有所不同,因为是对培养基进行检测而不破坏细胞本身。hGH瞬时检测系统适用于基因表达调控分析,在此用于研究猿猴病毒40增强子对单纯疱疹病毒胸苷激酶启动子的影响以及锌对小鼠金属硫蛋白-I启动子的影响。hGH的表达还可作为内部对照,与任何其他瞬时表达系统一起监测转染效率。迄今为止测试的所有细胞类型(包括AtT-20、CV-1、GC、GH4、JEG、L和原代垂体细胞)都能够将hGH分泌到培养基中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a38/367053/a76ab69f6720/molcellb00093-0163-a.jpg

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