Title: FLP mediated recombination of FRT sites in CHO-K1 cell line

Authors: E. Poluri, J.P. Gomedhikam, M.K. Kota, M. Kalagara, V.B. Bodanapu, B.P. Kota

Addresses: Department of Pharmaceutical Sciences, Pharmaceutical Biotechnology Division, Andhra University, Visakhapatnam 530 003, India. ' Department of Pharmaceutical Sciences, Pharmaceutical Biotechnology Division, Andhra University, Visakhapatnam 530 003, India. ' Department of Pharmaceutical Sciences, Pharmaceutical Biotechnology Division, Andhra University, Visakhapatnam 530 003, India. ' Department of Pharmaceutical Sciences, Pharmaceutical Biotechnology Division, Andhra University, Visakhapatnam 530 003, India. ' Department of Pharmaceutical Sciences, Pharmaceutical Biotechnology Division, Andhra University, Visakhapatnam 530 003, India ' Department of Pharmaceutical Sciences, Pharmaceutical Biotechnology Division, Andhra University, Visakhapatnam 530 003, India

Abstract: A binary system for gene activation and site specific integration based on the conditional recombination of transfected sequences mediated by the FLP recombinase from yeast was implemented in mammalian cells. In several cell lines, FLP rapidly and precisely recombined copies of its specific target sequences to activate an otherwise silent β-Galnreporter gene. Clones of marked cells were generated by excisional recombination within a chromosomally integrated copy of the silent reporters. These clones exhibited intense blue colour with X-Gal staining solution. The beta galactosidase activity expressed by these clones, were measured by cell lysis method.

Keywords: β-Gal; FLP recognition target site (FRT); pNEOβ-Gal; X-Gal.

DOI: 10.1504/IJBT.2004.004616

International Journal of Biotechnology, 2004 Vol.6 No.1, pp.94 - 100

Published online: 26 May 2004 *

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