Construction and Functional Analysis of Luciferase Reporter Plasmid Containing NF-H Gene Promoter

2014 ◽  
Vol 915-916 ◽  
pp. 942-946
Author(s):  
Tong Cun Zhang ◽  
Yao Meng ◽  
Nan Wang ◽  
Feng Lin ◽  
Tao Qin ◽  
...  

NF-H (a member of neurofilaments) is a protein widely expressed in all kinds of neurons after birth and can be used as one of the symbols of mature neurons. Constuction of NF-H promoter luciferase reporter plasmid will provide the theory basis for researching the effect of other transcription factors on regulating NF-H transcription. Here, human NF-H promoter luciferase reporter plasmid were successfully constructed. Then the effects of some key transcription factors were investigated by luciferase reporter assays in COS-7 cells. The results showed that retinoid X receptor α (RXRα) can enhance transcriptional activity of NF-H. Furthermore, ERK 1/2 (extracellular signal-regulated kinase 1/2) and STAT3 (signal transducer and activator of transcription 3) also show obvious impact in activating NF-H transcription.

2017 ◽  
Vol 8 ◽  
pp. 01007
Author(s):  
Hui Li ◽  
Jiapeng Li ◽  
Weilin Shi ◽  
Xiaoyu Zhang ◽  
Yuan Xiang ◽  
...  

2014 ◽  
Vol 556-562 ◽  
pp. 257-260
Author(s):  
Tong Cun Zhang ◽  
Yue Wang ◽  
Xing Hua Liao ◽  
Nan Wang ◽  
Hao Zhou

PCNA (proliferating cell nuclear antigen) is a protein related to tumor development, which has been used extensively in breast cancer diagnosis and prognosis. PCNA has proven to be a useful marker to evaluate cell proliferation and prognosis when combined with other breast cancer markers. Construction of PCNA promoter luciferase reporter plasmid will provide the theory basis for researching the effect of other transcription factors on regulating PCNA transcription. In this study, a human PCNA promoter luciferase reporter construct was generated by PCR amplification of PCNA promoter. The PCR fragment was digested and cloned into pGL3 vector. The promoter sequence was verified by sequencing. The results showed that luciferase reporter plasmids of PCNA promoter were successfully constructed. Then the effects of some key transcription factors, which play important roles in breast cancer cell proliferation, were investigated by luciferase reporter assays in MCF-7 cells. The results showed that ERα can enhance transcriptional activity of PCNA. Furthermore, 17-β-estradiol (E2) also shows an obvious impact in activating PCNA transcription. Our data illuminated that E2 enhances ERα-induced proliferation potential of MCF-7 cells by stimulating the transcriptional activity of PCNA. Our research will provide a model to screen some novel factors in regulating proliferation marker transcription.


2012 ◽  
Vol 424-425 ◽  
pp. 399-402
Author(s):  
Zhe Sun ◽  
Tong Cun Zhang ◽  
Yong Jiang

Aquaporins (AQPs), a family of water channels, function as a water-selective transporting protein in cell membranes. Aquaporin 1 (AQP1) was first discovered in human erythrocytes as a water channel for high osmotic water permeability. AQP1 promoter is responsible for regulation of its transcription and expression. In this study, molecular biology and genetic engineering knowledge and principles of construction of AQP1 promoter plasmid, and recombinant plasmids were identified and measured for luciferase, for promoter function. Our study successfully constructed AQP1 promoter luciferase reporter gene expression plasmid and successfully carry out the functions of its detection, provides a new foundation for the transcription expression and function regulation of the AQP1


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