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

AUTOR(ES)

Selden, R F

RESUMO

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.

Documentos Relacionados

• Expression of a human placental alkaline phosphatase gene in transfected cells: use as a reporter for studies of gene expression.
• The developmental regulation of the human zeta-globin gene in transgenic mice employing beta-galactosidase as a reporter gene.
• A reporter gene construct for studying the regulation of manganese peroxidase gene expression.
• Use of genetic recombination as a reporter of gene expression.
• Transient paralysis by heat shock of hormonal regulation of gene expression.