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Gulhane Med J. 2009; 51(2): 091-093


Expression of HBeAg gene region in Saccharomyces cerevisiae

Mehmet Yapar1, Kenan Şener1, Orhan Bedir2, Ertan Altaylı3, Ayhan Kubar1.

Abstract
'HBeAg/anti-HBe seroconversion is one of the markers frequently used in the diagnosis of hepatitis B infection. In this study, the “e” antigen gene region of the hepatitis B virus (HBV) was transformed to Saccharomyces cerevisiae and expressed by the yeast in order to obtain HBeAg protein used in serologic tests. Firstly, HBeAg gene region was amplified by PCR method. Later, the amplicons were cloned into the plasmid pYES2.1 via the TOPO® TA expression kit and transformed into the competent bacteria (TOPO10F' Escherichia coli). The plasmid pYES2.1+HBeAg was isolated from the competent bacteria, which was cultivated on LB media supplemented with ampicillin, and transformed to Saccharomyces cerevisiae via the S.c. Easy- Comp Transformation Kit. Finally, whether HBeAg was expressed by the yeast was checked through two different automatized systems. HbeAg producing yeast cells obtained may render it possible to purely isolate proteins in future studies.'

Key words: Gene expression, HBeAg, cloning



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