| Title |
An efficient rapid system for assaying HBx-mediated transactivation
|
|---|---|
| Published in |
Biotechnology Techniques, April 2017
|
| DOI | 10.1007/s10529-017-2334-1 |
| Pubmed ID | |
| Authors |
Xiaoling Zhou, Haijun Shi, Shaozhe Yang, Pingnan Sun |
| Abstract |
To develop a rapid and accurate assay system for screening inhibitors or enhancing agents targeting the transactivation capability of hepatitis B virus X protein (HBx) that activates cellular promoters in host cells to facilitate viral replication. We constructed a new GFP-based reporter system which was different from a luciferase-based reporter system. Firstly, a FLAG-tagged HBx gene was inserted into an expression plasmid, resulting in plasmid pHBx. Next, HBx-FLAG was linked to EGFP by the internal ribosome entry site resulting in plasmid pHBxE. The transactivation effect of HBx-flag on cytomegalovirus (CMV) promoter was verified by EGFP expression using fluorescence quantitation and qPCR. Furthermore, the transactivation ability of the HBx gene was quantified by flow cytometry. Finally, this assay system was tested by known regulators of HBx including DDB1, ID1, and P53. As expected, the GFP reporter level in 293T cells changed with the increasing of HBx regulators. Furthermore, the system modeling the function of transactivation repressor in Hep3B, a HBV-integrated cell line. Collectively, the GFP-based reporter system provides a rapid and accurate approach for analyzing transactivation ability of HBx. |
X Demographics
The data shown below were collected from the profile of 1 X user who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 5 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 5 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Librarian | 1 | 20% |
| Student > Ph. D. Student | 1 | 20% |
| Researcher | 1 | 20% |
| Unknown | 2 | 40% |
| Readers by discipline | Count | As % |
|---|---|---|
| Arts and Humanities | 1 | 20% |
| Immunology and Microbiology | 1 | 20% |
| Medicine and Dentistry | 1 | 20% |
| Unknown | 2 | 40% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 08 April 2017.
All research outputs
#22,764,772
of 25,382,440 outputs
Outputs from Biotechnology Techniques
#2,493
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Outputs of similar age
#284,669
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Outputs of similar age from Biotechnology Techniques
#23
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