USF2与其截短体真核表达载体的构建及对Smurf1/2转录水平的影响

山东大学学报(医学版) ›› 2013, Vol. 51 ›› Issue (8): 34-37.

USF2与其截短体真核表达载体的构建及对Smurf1/2转录水平的影响

谭雅文，高海东

山东大学齐鲁医院乳腺外科，济南 250012

收稿日期:2012-12-30 出版日期:2013-08-10 发布日期:2013-08-10
通讯作者: 高海东， E-mail：oncology@sdu.edu.cn

Construction of the eukaryotic expression vectors of USF2 gene and its truncated genes and their influence on Smurf1/2 transcriptional level

TAN Ya-wen, GAO Hai-dong

Department of Breast Surgery, Qilu Hospital of Shandong University, Jinan 250012, China

Received:2012-12-30 Online:2013-08-10 Published:2013-08-10

摘要/Abstract

摘要：

目的构建上游刺激因子2(USF2)及其截短体的真核表达载体，鉴定USF2蛋白中抑制泛素连接酶Smurf1/2转录的功能区域。方法采用PCR技术扩增USF2 及其两个截短体USF2(1~235aa)、USF2(236~346aa)编码基因，分别将其构建在pCMV-Myc重组载体上，转染真核细胞后验证其表达，Western blot及实时定量PCR确定USF2下调Smurf1/2转录水平的区域。结果成功地将USF2及其两个截短体编码基因构建至pCMV-Myc载体，并验证其在真核细胞中的正确表达，Western Blot和实时定量PCR实验结果显示，USF2 C端236~346aa区域对Smurf1/2的转录水平有抑制效应。结论 USF2通过其C端236~346aa区域对Smurf1/2的转录产生抑制效应。

关键词: 上游刺激因子2；Smad泛素调节因子1/2；转录水平；截短体

Abstract:

Objective To construct the eukaryotic expression vectors of Upstream Stimulatory Factor 2 (USF2) and its truncated genes，and identify the functional region of USF2 protein down regulating the Smurf1/2 transcriptional level. Methods Using polymerase chain reaction and gene recombination techniques，USF2 gene and its two truncated genes USF2(1-235aa) and USF2(236-346aa) were amplified and cloned into the pCMV-Myc vector respectively, and then transfected into MCF7 cells to detect their expressions. The 236-346aa region at the C terminal of USF2 inhibiting the Smurf1/2 transcription was explored by Western blot and real-time PCR techniques. Results USF2 and its truncated genes were successfully cloned into the pCMV-Myc vector. Through Western blot and real-time PCR, the 236-346aa region at the C terminal of USF2 inhibiting the transcriptional level of Smurf1/2 was proved. Conclusion USF2 suppresses the Smurf1/2 transcriptional level through the 236-346aa region at the C terminal．

Key words: Upstream transcription factor 2; Smad ubiquitination regulator factor 1/2; Transcriptional level； Truncated genes

中图分类号:

谭雅文，高海东. USF2与其截短体真核表达载体的构建及对Smurf1/2转录水平的影响[J]. 山东大学学报(医学版), 2013, 51(8): 34-37.

TAN Ya-wen, GAO Hai-dong . Construction of the eukaryotic expression vectors of USF2 gene and its truncated genes and their influence on Smurf1/2 transcriptional level[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(8): 34-37.

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