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浙江大学学报（医学版）
Journal of ZheJiang University(Medical Science)  2011, Vol. 40 Issue (4): 427-431    DOI: 10.3785/j.issn.1008-9292.2011.04.014
    
Establishment of NCAM L1 minigene model and its splicing patterns in different cell lines
ZHANG Wei1 ,SHEN Quan 1, PENG Zheng-Yu 2
1.School of Medicine and Life Science,Zhejiang University City College,Hangzhou 310015,China; 2.Institute of Biomedical Sciences,Fudan University,Shanghai 200032,China
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Abstract  Objective： To establish a minigene model of neural cell adhesion molecule L1 (NCAM L1)gene and to study its aplicing patterns in different cell lines．
Methods： Using human genetic cDNA as template,the NCAM L1 minigene fragment was amplified and inserted into eukaryotic expression vector. The minigene was transfected into 4 cell lines and the splicing patterns of NCAM L1 minigene in these cell lines were studied．
Results： The splicing patterns of NCAM L1 minigene were different in individual cell lines．In PFSK and Hela cell lines,two splicied isoforms were generated but in COS-1 and R28 cell lines,only one isoform existed.
Conclusion： NCAM L1 minigene model can be used in alternative splicing analysis.

Key words： Hela cells      Plasmids      Genetic vectors      Spliceosomes      NCAM L1； Mini-gene； Pre-mRNA； Alternative splicing     
Published: 25 July 2010
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Cite this article:

. Establishment of NCAM L1 minigene model and its splicing patterns in different cell lines. Journal of ZheJiang University(Medical Science), 2011, 40(4): 427-431.

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https://www.zjujournals.com/med/10.3785/j.issn.1008-9292.2011.04.014     OR     https://www.zjujournals.com/med/Y2011/V40/I4/427


NCAM L1小基因模型的建立及其剪接模式研究

目的：建立可用于前体mRNA可变剪接分析的NCAM L1小基因模型。
方法：以人基因组DNA为模板,通过PCR 扩增获得NCAM L1小基因片段,并将其克隆至真核表达载体中,构建小基因的质粒。在此基础上,用NCAM L1小基因模型转染HeLa、COS-1、PFSK及R28细胞,并用RT-PCR进行被剪接的小基因产物的半定量检测。
结果：NCAM L1小基因在4种细胞中的剪接模式不同，在PFSK以及Hela细胞中，存在2种剪接亚型，而在COS-1以及R28细胞中只有一种剪接亚型存在。
结论：所构建的NCAM L1小基因可用于细胞水平的基因剪接分析。

关键词： Hela细胞,  质粒,  遗传载体,  剪接体,  NCAM L1,  小基因,   ,  前体mRNA,   ,  可变剪接 
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