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 

机构地区：[1]Laboratory of Genomic Physiology, School of Life Sciences, Fudan University, Shanghai 200433, China

出　　处：《Science China(Life Sciences)》2004年第3期211-218,共8页中国科学(生命科学英文版)

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.

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.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.

关 键 词：MINIGENE GluR-B FGF-2R ZIS Tra2β pre-mRNA splicing. 

分 类 号：N[自然科学总论]