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Cloning and functional analysis of the promoter of ZNF268 gene.

Xiao Peng, Yan Sun, Hui Liu, Deming Gou, Wenxin Li

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Abstract

The human ZNF268 gene is a novel C2H2 zinc finger gene restrictively expressed in the liver of human embryo. As an initial study on the regulation of its expression, 2.5 kb fragment of the ZNF268 5'-flanking region was cloned from human genome by PCR. This fragment was inserted into pEGFP1 vector and then the recombinant plasmid was transfected into several cell lines by liposomal transfection method. EGFP expression was observed in four cell lines under confocal laser scanning microscope. A series of 5'-deletion fragments from -2456(-)+77 bp to -20(-)+77 bp were inserted into pCAT-Basic vector to construct recombinant reporter plasmids, which were then transfected into Hela cells for deletion analysis. Results of deletion analysis revealed that an important region for transcriptional activity lies between -156 and -20 bp.

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What this paper is about

The human ZNF268 gene is a novel C2H2 zinc finger gene restrictively expressed in the liver of human embryo. As an initial study on the regulation of its expression, 2.5 kb fragment of the ZNF268 5'-flanking region was cloned from human genome by PCR. This fragment was inserted into pEGFP1 vector and then the recombinant plasmid was transfected into several cell lines by liposomal transfection method. EGFP expression was observed in four cell lines under confocal laser scanning microscope. A series of 5'-deletion fragments from -2456(-)+77 bp to -20(-)+77 bp were inserted into pCAT-Basic vector to construct recombinant reporter plasmids, which were then transfected into Hela cells for deletion analysis. Results of deletion analysis revealed that an important region for transcriptional activity lies between -156 and -20 bp.

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Available abstract

The human ZNF268 gene is a novel C2H2 zinc finger gene restrictively expressed in the liver of human embryo. As an initial study on the regulation of its expression, 2.5 kb fragment of the ZNF268 5'-flanking region was cloned from human genome by PCR. This fragment was inserted into pEGFP1 vector and then the recombinant plasmid was transfected into several cell lines by liposomal transfection method. EGFP expression was observed in four cell lines under confocal laser scanning microscope. A series of 5'-deletion fragments from -2456(-)+77 bp to -20(-)+77 bp were inserted into pCAT-Basic vector to construct recombinant reporter plasmids, which were then transfected into Hela cells for deletion analysis. Results of deletion analysis revealed that an important region for transcriptional activity lies between -156 and -20 bp.

Key concepts: Biology, Transfection, Molecular biology, Plasmid, Cloning (programming), Gene, Cloning vector, Recombinant DNA

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