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 Tra2beta1 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.
Li
Jing
J
Laboratory of Genomic Physiology, School of Life Sciences, Fudan University, Shanghai 200433, China.
Chen
Xianhua
X
Lin
Wanmin
W
Li
Lishu
L
Han
Yu
Y
Xu
Ping
P
D017398:Alternative Splicing / Q000235:genetics*; D000818:Animals; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008957:Models, Genetic; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics; D051497:Receptor, Fibroblast Growth Factor, Type 2; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics; D017470:Receptors, Glutamate / Q000235:genetics
D020033:Protein Isoforms; D012322:RNA Precursors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D017468:Receptors, Fibroblast Growth Factor; D017470:Receptors, Glutamate; C486374:Zranb2 protein, mouse; C496353:FGFR2 protein, human; C496354:Fgfr2 protein, mouse; D020794:Receptor Protein-Tyrosine Kinases; D051497:Receptor, Fibroblast Growth Factor, Type 2