Plasmid Construction of TMEM16A-pcDNA3.1 and its Application to Transient and Stable Transfection of FRT Cells

Feng Hao; Zhong Hai Yuan; Zhi Xin Wang; Hui Jing Xu; Fang Fang; Xin Gang Guan; Jiang Yong; Li Yan

doi:10.4028/www.scientific.net/AMR.554-556.1734

Paper Titles

The Study Evolution of TB in Black Tea
p.1717

The Immobilization of VEGF-Fc for Promoting Endothelial Cells Proliferation in Porous PCL Scaffolds
p.1721

The Establishing and Applied Research of Animal Models of Mammary Precancerous Lesions
p.1725

Comparison of Chloroplast TrnL-F Sequences of 5 Common Potentilla Species in Jilin Province of China
p.1730

Plasmid Construction of TMEM16A-pcDNA3.1 and its Application to Transient and Stable Transfection of FRT Cells
p.1734

Comparison of Urinary Crystallites from Patients with Renal Calculi with that from Healthy Subjects
p.1738

Metabonomic Investigation on Rats’ Dynamic Responses to Cantonese Herbal Tea Intake
p.1742

Vector-Based siRNA is a Powerful Tool to Reduce Protein Expression in Cancer Research
p.1747

Study on Biological Safety of TiO₂ Nanomaterials
p.1751

HomeAdvanced Materials ResearchAdvanced Materials Research Vols. 554-556Plasmid Construction of TMEM16A-pcDNA3.1 and its...

Plasmid Construction of TMEM16A-pcDNA3.1 and its Application to Transient and Stable Transfection of FRT Cells

Abstract:

Calcium-activated chloride channels (CaCCs) play pivotal roles in many physiological Activities, including transepithelial fluid secretion, smooth muscle contraction and sensory transduction. TMEM16A is a bona fide calcium-activated chloride channel，which was discovered by three independent labs in 2008 after Calcium-activated chloride channel current was recorded about thirty years ago. In this study, DNA fragments encoding mouse TMEM16A with green fluorescence protein (GFP) fusion protein were subcloned into pcDNA3.1/Zeo. Transient transfection condition was optimized and Fischer Thyroid epithelial cells (FRT) expressing TMEM16A were got by stable transfection. The classical calcium-activated chloride channels current was recorded in FRT cells stably expressing TMEM16A by whole cell patch clamp technique. These results were beneficial for the delving into the effects of other bivalent cations on TMEM16A-CaCCs and the role of TMEM16A-CaCCs in cell proliferation and migration.

You might also be interested in these eBooks

View Preview

Info:

Periodical:

Advanced Materials Research (Volumes 554-556)

Pages:

1734-1737

DOI:

https://doi.org/10.4028/www.scientific.net/AMR.554-556.1734

Citation:

Cite this paper

Online since:

July 2012

Authors:

Feng Hao*, Zhong Hai Yuan, Zhi Xin Wang, Hui Jing Xu, Fang Fang, Xin Gang Guan, Jiang Yong, Li Yan

Keywords:

CaCCs, Eukaryotic Expression Vector, Patch Clamp, Stable Transfection, TMEM16A

Export:

RIS, BibTeX

Price:

Permissions CCC:

Request Permissions

Permissions PLS:

Request Permissions

Сopyright:

Citation:

* - Corresponding Author

References

[1] Yang YD, Cho H, Koo JY, Tak MH, Cho Y, Shim WS, Park SP, Lee J, Lee B, Kim BM, Raouf R, Shin YK, Oh U. Nature. 2008 Oct 30;455(7217):1210-5.

DOI: 10.1038/nature07313

Google Scholar

[2] Caputo A, Caci E, Ferrera L, Pedemonte N, Barsanti C, Sondo E, Pfeffer U, Ravazzolo R, Zegarra-Moran O, Galietta LJ. Science. 2008 Oct 24;322(5901):590-4.

DOI: 10.1126/science.1163518

Google Scholar

[3] Schroeder, B. C., Cheng, T., Jan, Y. N. & Jan, L. Y. Cell. 2008; 134: 1019–1029.

Google Scholar

[4] HAO Feng, YI Fei, ZHANG Di, NING Yan, SU Wei-heng, FENG Xue-chao, YANG Hong, MA Tong-hui. Chem. Res. Chinese U. 2011, 27(3) :461-3.

Google Scholar

[5] Duran C, Hartzell HC.Acta Pharmacol Sin. 2011 Jun;32(6):685-92.

Google Scholar

[6] Hartzell HC, Yu K, Xiao Q, Chien LT, Qu Z. J Physiol. 2009 May 15;587(Pt 10):2127-39.

Google Scholar

[7] Miettinen M, Wang ZF, Lasota J. Am J Surg Pathol. 2009 Sep;33(9):1401-8.

Google Scholar