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

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 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.

scholarly journals Identification of Serum miRNA-423-5p Expression Signature in Somatotroph Adenomas

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Sida Zhao ◽  
Jianhua Li ◽  
Jie Feng ◽  
Zhenye Li ◽  
Qian Liu ◽  
...  
Keyword(s):  
Expression Profiling ◽  
Gh3 Cells ◽  
Luciferase Reporter ◽  
Reporter Plasmid ◽  
Circulating Mirnas ◽  
Luciferase Reporter Gene ◽  
Luciferase Reporter Plasmid ◽  
Exosomal Mirnas ◽  
Gene Assay ◽  
Mirnas Expression

Circulating miRNAs are novel disease biomarkers that are valuable for diagnosis and prognosis. But the circulating miRNAs profile in somatotroph adenomas is still unknown. Therefore, serum exosomal miRNAs expression profiling in somatotroph adenomas was performed on 6 somatotroph adenomas and 6 normal controls. From the exosomal miRNAs expression profiling, we found 169 miRNAs differently expressed between somatotroph adenomas and healthy pituitary samples (p< 0.05, FC > 2). Among the 169 miRNAs, miR-423-5p was expressed lower in somatotroph adenomas than in healthy pituitary samples, which was proved by miRSCan Panel Chip™ qPCR. PTTG1 and SYT1 were the target mRNAs of miR-423-5p, and transcriptomics and proteomics profile both indicated the high expression of PTTG1 and SYT1 in somatotroph adenomas. H-scores were 223.1 ± 34.7 for PTTG1 and 163.4 ± 42.3 for SYT1 in 62 somatotroph adenomas specimens and 84.2 ± 21.3 for PTTG1 and 47.4 ± 17.2 for SYT1 in 6 healthy pituitary specimens by IHC. miR-423-5p inhibited the expression of SYT1 and PTTG1 at the mRNA and protein levels. Dual luciferase reporter gene assay shown was significantly reduced in the presence of miR-423-5p in GH3 cells transfected with wild-type PTTG1 3'UTR luciferase reporter plasmid but not reduced when transfected with the mutation PTTG1 3'UTR luciferase reporter plasmid (p<0.01). In vitro experiments showed that miR-423-5p induced cell apoptosis, inhibited cell proliferation, and reduced growth hormone release and migration of GH3 cells. The activity of miR-423-5p in GH3 cell was nearly blocked by its inhibitor. These results verified the central role of low miR-423-5p in promoting tumorigenesis in somatotroph adenomas. PTTG1 may act as biomarkers for clinical treatment of somatotroph adenomas.

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.

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