Establishment and application of minigene models for studying pre-mRNA alternative splicing

The objective of the present study is to establish a minigene model for studying pre-mRNA alternative splicing. To prepare the minigene DNA constructs, with human or mouse genomic DNA as templates, GluR-B, FGF-2R and Zis “minigene” fragments were amplified using PCR and cloned to the eukaryotic expression vectors. The three constructed minigenes and the expression vectors of Tra2β1 and Zis2 were co-transfected in Hela cells. RT-PCR analysis was performed to semi-quantitatively determine the spliced products from the minigenes. The results demonstrated that the constructed minigenes are useful in studying the pre-mRNA alternative splicing in cultured cells. With the established Zis minigene, we for the first time found that Zis2 isoform regulates the alternative splicing of Zis minigene.     

Red cell ouabain binding sites, Na+K+-ATPase, and intracellular Na+ as individual characteristics

Healthy male volunteers were infused for three hours with either a dopamine hydrochloride solution at a rate of 4 ug/kg/min or with normal saline. Plasma amine oxidase and platelet MAO activity towards benzylamine both increased in response to intravenous dopamine. There was no increase in enzyme activity when dopamine was added to the platelet and plasma enzymes in vitro. This heretofore unreported increase in the oxidative deaminating capacity of the human organism may represent an adaptive physiologic response to the high circulating levels of dopamine and provides further evidence for a possible functional significance of these enzymes in man.