Establishment and Characterization of Stably Transfected MDCK Cell Line, Expressing Hbest1 Protein

K. MLADENOVA1, V. MOSKOVA-DOUMANOVA1, I. TABASHKA2, S. PETROVA2, Z. LALCHEV2 and J. DOUMANOV2
1 Sofia University “St. Kliment Ohridski”, Department of Cytology, Histology and Embryology, Faculty of Biology, BG – 1164 Sofia, Bulgaria
2 Sofia University “St. Kliment Ohridski”, Department of Biochemistry, Faculty of Biology, BG – 1164 Sofia, Bulgaria

Abstract

Mladenova, K., V. Moskova-Doumanova, I. Tabashka, S. Petrova, Z. Lalchev and J. Doumanov, 2013. Establishment and characterization of stably transfected MDCK cell line, expressing hBest1 protein. Bulg. J. Agric. Sci., Supplement 2, 19: 159–162

 

Bestrophin-1(Best1) is a transmembrane protein specifically expressed in the retinal pigment epithelial cells. It is localized on their basolateral plasma membrane. Malfunctions of the protein lead to retinal pathologies, named Bestrophinophaties. Best1 is thought to be a Ca2+ - activated Cl channel or a regulator of ion transport, or both. Some other functions in the maintaining of cellular homeostasis are also attributed to this protein. In order to obtain additional information about protein structure and functions we established a stably transfected MDCK II cell line expressing human Best1, conjugated with Myc and His- tags at C-terminus. The presence of stably expressed Best1 protein was demonstrated by immunofluoresce studies. Confocal microscopy images confirmed its basolateral localization. Additionally, no differences in the cell growth ratio and mitotic index of established cell line, in comparison with conventional MDCK cells were observed. Our studies suggest that Best1 does not influence cell growth and cell polarity in transfected cells, making our stably transfected cell line an appropriate model for investigations of Best1 functions.

Key words: Best1 protein, BVMD, MDCK II cells

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