Function Analysis of Luciferase Reporter Plasmid with AQP1 Gene Promoter

2011 ◽  
Vol 396-398 ◽  
pp. 1486-1489
Author(s):  
Yong Jiang ◽  
Zhe Sun ◽  
Tong Cun Zhang
Keyword(s):  
Transcriptional Regulation ◽  
Function Analysis ◽  
Gene Promoter ◽  
Luciferase Reporter ◽  
Reporter Plasmid ◽  
Reporter Construct ◽  
Luciferase Reporter Construct ◽  
Transcription Activity ◽  
Luciferase Reporter Plasmid ◽  
Precise Role

The aquaporins (AQP) are a family of homologous water transporting proteins that are expressed in many epithelial, endothelial and other tissues. Myocardin is important for SMC differentiation, but its precise role in regulating the initiation of AQP1 transcription activity is less clear. Function analysis of AQP1 promoter luciferase reporter plasmid will provide the theory basis for researching the function of transcription activity. In this study, the rat and human myocardin promoter luciferase reporter construct were successfully constructed. Then to determine whether AQP1 transcription activity were regulated by myocardin, luciferase repoeter assays were performed in COS-7 cells. The results illustrated that myocardin significantly activated rat and human AQP1 promoter. AQP1 promoter transactivity was inhibited by △Q. The present study provided the first evidence that myocardin may have an influence on the expression of AQP1 and reveal a basis of the mechanism transcriptional regulation of AQP1.

scholarly journals MKL1 and STAT3 activate the activity of the luciferase reporter plasmid containing the CAAP1 gene promoter

2019 ◽  
Vol 78 ◽  
pp. 01003
Author(s):  
Jun-Yan Li ◽  
Zhu Yu ◽  
Feng-Yun Wang
Keyword(s):  
Breast Cancer ◽  
Breast Cancer Cells ◽  
Gene Promoter ◽  
Luciferase Reporter ◽  
Reporter Assay ◽  
Reporter Plasmid ◽  
Transcription Activity ◽  
Luciferase Reporter Plasmid ◽  
The Relationship ◽  
Mcf 7

Breast cancer is the leading cause of cancer death in women worldwide. The etiology of the disease is not yet clear. We know that MKL1 and STAT3 play an important part in the development and progression of breast cancer. CAAP1 is a ubiquitous and highly conserved protein that is closely related to the apoptotic process of tumors. However, the definitive transcriptional mechanism of the CAAP1 gene is still unclear. In our study, we constructed a luciferase reporter plasmid for the human CAAP1 gene promoter. Then one or both of the two overexpression vectors of MKL-1 and STAT3 were co-transfected into MCF-7 cells with CAAP1 promoter plasmid, and we then tested activation of the CAAP1 promoter by luciferase reporter assay. The results show that compared with the transfected pcDNA3.1 group, MKL1 can evidently increase the transcription activity of the CAAP1 gene promoter, while the STAT3 group can slightly upregulate the transcription activity of the CAAP1 gene promoter. Our research will further reveal the relationship between CAAP1 and the occurrence and development of breast cancer cells, and provide a new idea and direction for the cures of breast cancer.

scholarly journals Construction and functional analysis of luciferase reporter plasmid containing connexin43 gene promoter

2017 ◽  
Vol 8 ◽  
pp. 01007
Author(s):  
Hui Li ◽  
Jiapeng Li ◽  
Weilin Shi ◽  
Xiaoyu Zhang ◽  
Yuan Xiang ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Gene Promoter ◽  
Luciferase Reporter ◽  
Reporter Plasmid ◽  
Luciferase Reporter Plasmid

Construction and Functional Analysis of Luciferase Reporter Plasmid Containing PCNA Gene Promoter

2014 ◽  
Vol 556-562 ◽  
pp. 257-260
Author(s):  
Tong Cun Zhang ◽  
Yue Wang ◽  
Xing Hua Liao ◽  
Nan Wang ◽  
Hao Zhou
Keyword(s):  
Breast Cancer ◽  
Cell Proliferation ◽  
Transcription Factors ◽  
Transcriptional Activity ◽  
Nuclear Antigen ◽  
Luciferase Reporter ◽  
Reporter Plasmid ◽  
Luciferase Reporter Construct ◽  
Luciferase Reporter Plasmid ◽  
Mcf 7

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.

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 ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Gene Promoter ◽  
Luciferase Reporter ◽  
Reporter Plasmid ◽  
Extracellular Signal Regulated Kinase ◽  
Luciferase Reporter Plasmid ◽  
Extracellular Signal ◽  
Mature Neurons ◽  
Reporter Assays ◽  
Transcription 3

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.

Construction of Luciferase Reporter Plasmid with Human AQP1 Gene Promoter

2012 ◽  
Vol 424-425 ◽  
pp. 399-402
Author(s):  
Zhe Sun ◽  
Tong Cun Zhang ◽  
Yong Jiang
Keyword(s):  
Water Channel ◽  
Gene Promoter ◽  
Luciferase Reporter ◽  
Reporter Plasmid ◽  
Luciferase Reporter Gene ◽  
Expression Plasmid ◽  
Luciferase Reporter Plasmid ◽  
Osmotic Water ◽  
And Function ◽  
Transcription Expression

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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