尹安琪 孙佳辰 张娜娜 药立波 李燕 熊利泽.NDRG2 基因六种可变剪接体的分型鉴定[J].现代生物医学进展英文版,2016,16(20):3835-3838. |
NDRG2 基因六种可变剪接体的分型鉴定 |
Identification of Alternatively Spliced Variants of Human NDRG2 Gene and its Application |
|
DOI: |
中文关键词: N-myc 下游调节基因2 剪接体 基因表达 PCR |
英文关键词: NDRG2 Splice variant Gene expression PCR |
基金项目:国家自然科学基金项目(81471110) |
|
Hits: 935 |
Download times: 0 |
中文摘要: |
目的:抑癌基因N-myc 下游调节基因2(N-myc down streamregulated gene 2, NDRG2)在抑制肿瘤的发生及进展过程中具有
重要作用,本研究旨在区分并鉴定NDRG2 基因mRNA 的6 种可变剪接体。方法:针对NDRG2 基因mRNA6 种剪接体外显子的
差别设计6 对特异性引物。分别从正常人脑组织和胶质瘤细胞U251 中提取总RNA,反转录为cDNA,利用高保真酶扩增目的剪
接体DNA,并根据PCR扩增产物电泳条带的有无和大小初步判断剪接体的型别,最后将PCR产物胶回收进行测序鉴定。结果:
利用本实验中设计的引物可以将人脑组织和U251 胶质瘤细胞中的NDRG2 基因mRNA剪接体经行分型鉴定,确定了分型引物
的可行性。人脑组织和U251 细胞中均含有A、B、D 和E 四种剪接体。结论:利用本研究设计的特异性引物进行PCR扩增,可以判
断某种细胞或组织中表达的NDRG2 mRNA分子剪接体的型别。有助于未来深入研究抑癌基因NDRG2 在肿瘤发生发展中的抑
制作用与不同剪接体的表达相关性。 |
英文摘要: |
Objective:Tumor suppressor gene N-myc 2(N-myc down stream regulated gene 2, NDRG2)plays important role in
suppressing the tumorigenesis and progressing. Our research aims to differentiate and identify six variants of NDRG2 mRNA.Methods:Based on the differences of six variants, we designed six specific primers. We extracted total RNA from normal brain tissue and glioma
cell line U251 respectively, using RT-PCR to get the targeted cDNA. Then we differentiate the six variants by electrophoresis. Finally we
recollected the RT-PCR products to test the sequencing.Results:With the specific primers we confirmed that both normal human brain
tissue and glioma cell line U251 contain the A, B, D, and E, four kinds of variants.Conclusion:With the specific primers we designed,
we can tell the differences of NDRG2 variants expression in different cells or tissues, which help to further explore the relationship
between the NDRG2 variants expression and the tumor suppressing effect. |
View Full Text
View/Add Comment Download reader |
Close |