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

作     者：LI Jing CHEN Xianhua LIN Wanmin LI Lishu HAN Yu XU Ping 

作者机构：Laboratory of Genomic Physiology School of Life Sciences Fudan University Shanghai 200433 China 

出 版 物：《Science China(Life Sciences)》 (中国科学(生命科学英文版))

年 卷 期：2004年第47卷第3期

页      面：211-218页

核心收录：

学科分类：07[理学] 08[工学] 

基　　金：This work was supported by the National Natural Science Foundation of China(Grant No.30070242) Scientific Foundation for Youth in Life Sciences,Fudan University 

主　　题：minigene GluR-B FGF-2R Zis Tra2β pre-mRNA splicing. 

摘      要：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 搈inigene?fragments were amplified us-ing PCR and cloned to the eukaryotic expression vectors. The three constructed minigenes and the expression vectors of Tra2b1 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 al-ternative 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.