Establishment and application of minigene models for studying pre-mRNA alternative splicing |
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Authors: | LI Jing CHEN Xianhua LIN Wanmin LI Lishu HAN Yu XU Ping |
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Institution: | Laboratory of Genomic Physiology, School of Life Sciences, Fudan University, Shanghai 200433, China. |
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Abstract: | 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. |
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Keywords: | minigene GluR-B FGF-2R Zis Tra2b pre-mRNA splicing |
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