Expression and activity analysis of mouse mdm2

WANG Xiao-du; ZHAO A-yong; DENG Xu-fang; MA Zhi-yong

doi:10.11833/j.issn.2095-0756.2012.02.001

Volume 29 Issue 2

Mar. 2012

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WANG Xiao-du, ZHAO A-yong, DENG Xu-fang, MA Zhi-yong. Expression and activity analysis of mouse mdm2[J]. Journal of Zhejiang A&F University, 2012, 29(2): 155-160. doi: 10.11833/j.issn.2095-0756.2012.02.001

Citation:

WANG Xiao-du, ZHAO A-yong, DENG Xu-fang, MA Zhi-yong. Expression and activity analysis of mouse mdm2[J]. Journal of Zhejiang A&F University, 2012, 29(2): 155-160. doi: 10.11833/j.issn.2095-0756.2012.02.001

Expression and activity analysis of mouse mdm2

doi: 10.11833/j.issn.2095-0756.2012.02.001

1.
School of Forestry and Biotechnology，Zhejiang A & F University，Lin’an 311300，Zhejiang，China;
2.
Shanghai Veterinary Research Institute，Chinese Academy of Agricultural Science，Shanghai 200241，China

More Information

Corresponding author: MA Zhi-yong
Received Date: 2011-07-20
Rev Recd Date: 2011-10-26
Publish Date: 2012-04-20

Abstract

The mdm2 （murine double minute 2） is one of the factors inhibiting p53 activity，and their interaction is important to hold cell survival. In this paper，mouse mdm2 gene was cloned from mouse 3T3 cell strain，and the recombinant plasmid p3xFLAG-CMV-7.1-mdm2 was constructed and confirmed by restriction enzyme digestion and DNA sequencing. Western blotting analysis and indirect immune-fluorescent analysis showed that FLAG-mdm2 expressed in p3xFLAG-CMV-7.1-mdm2-transfected H1299 cells，which was 60 kDa in size and dominantly localized in nucleus. The activity of FLAG-mdm2 protein was confirmed by p53 ubiquitination assay and luciferase assay. The result showed that mdm2 cell model inhibiting p53 activity is established. Our results would be useful for studying the mdm2 biological fuction in regulating p53 activity and interaction between mdm2 and p53.［Ch，5 fig. 15 ref.］
- zoology,
- mdm2 expression,
- mdm2 acitivity,
- cell model

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Expression and activity analysis of mouse mdm2

1. School of Forestry and Biotechnology，Zhejiang A & F University，Lin’an 311300，Zhejiang，China;

2. Shanghai Veterinary Research Institute，Chinese Academy of Agricultural Science，Shanghai 200241，China

Corresponding author: MA Zhi-yong

Received Date: 2011-07-20

Rev Recd Date: 2011-10-26

Publish Date: 2012-04-20

Key words:

Abstract: The mdm2 （murine double minute 2） is one of the factors inhibiting p53 activity，and their interaction is important to hold cell survival. In this paper，mouse mdm2 gene was cloned from mouse 3T3 cell strain，and the recombinant plasmid p3xFLAG-CMV-7.1-mdm2 was constructed and confirmed by restriction enzyme digestion and DNA sequencing. Western blotting analysis and indirect immune-fluorescent analysis showed that FLAG-mdm2 expressed in p3xFLAG-CMV-7.1-mdm2-transfected H1299 cells，which was 60 kDa in size and dominantly localized in nucleus. The activity of FLAG-mdm2 protein was confirmed by p53 ubiquitination assay and luciferase assay. The result showed that mdm2 cell model inhibiting p53 activity is established. Our results would be useful for studying the mdm2 biological fuction in regulating p53 activity and interaction between mdm2 and p53.［Ch，5 fig. 15 ref.］

Expression and activity analysis of mouse mdm2

doi: 10.11833/j.issn.2095-0756.2012.02.001

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

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