alexa Abstract | Establishment of EGFP-Purα Fusion Fluorescent Protein Cell Lines And Stable Expression of Purα Protects Cells From DNA Damage

Journal of Neurology and Neuroscience
Open Access

OMICS International organises 3000+ Global Conferenceseries Events every year across USA, Europe & Asia with support from 1000 more scientific Societies and Publishes 700+ Open Access Journals which contains over 50000 eminent personalities, reputed scientists as editorial board members.

Open Access Journals gaining more Readers and Citations

700 Journals and 15,000,000 Readers Each Journal is getting 25,000+ Readers

This Readership is 10 times more when compared to other Subscription Journals (Source: Google Analytics)

Research Article Open Access


Purα is a multifunctional protein that plays an important roles in DNA repair and cell cycle regulation. Purα knockout mice have been successfully established but unfortunately the knockout mice die at the 4th week after birth and that impeded the functional research of Purα in nervous system. The cells originated from nervous system is difficult for the transfection with the routine approaches and the lower transfection efficiency also hindered the research of gene manipulation in the nervous system. The lentivirus provided a new thoroughfare for the research of gene function in nervous system. On the current paper we reported here that lentivirus vector with overexpression of EGFP-Purα fusion fluorescent protein and its control-lentivirus vector with EGFP have been successfully constructed. The constructs were packaged and the human glioblastoma cell, U87MG were infected with these two lentivirus and selected with puromycin to establish the stable cell lines with overexpression of EGFP-Purα fusion protein. The expression level in the established cell lines were examined with qPCR and western blotting assay, the results demonstrated that Purα can be stably expressed in the established cell lines. The further experiments were employed to detect the effects of Purα on cell growth and proliferation with cell counting assay (CCK-8), the results proved that when the cells were treated with different concentrations of HU, the rate of cell survival in Purα overexpression group is higher than that in the control group, it suggested that Purα could protect the cells from the toxicity of HU. The protective effects of Purα on DNA damage were also evaluated with examination of the changes of the amount of DNA damage associated protein, γH2AX, as well as the pulse field gel assay, all the results illustrated that when cells were treated with 2mM HU for 18 hours, the amount of γH2AX is much less in Purα overexpression cell line than that in the control group, pulse field gel assay also exhibited the less DNA damage in Purα overexpression group than that in the control groups. Above all, these results confirmed that Purα plays a protective role in the DNA damage induced by HU.

To read the full article Peer-reviewed Article PDF image | Peer-reviewed Full Article image

Author(s): Juan Chai Yongling Li Zhongfa Jia Chengmin Yuan Ping Li Tao Sun Jianguo Niu Huichen Wang Jianqi Cui


Purα, U87MG cell, γH2AX, stable cell lines, DNA damage, Translational Neuroscience,Clinical Neurology

Peer Reviewed Journals
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version