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  • Characterization of the 5'-flanking region of the gene encoding the 50 kDa subunit of human DNA polymerase delta.

Characterization of the 5'-flanking region of the gene encoding the 50 kDa subunit of human DNA polymerase delta.

Biochimica et biophysica acta (2000-09-09)
A Perez, A Leon, M Y Lee
ABSTRACT

DNA polymerase delta consists of at least four subunits: p125, p68, p50, and p12 [Liu et al., J. Biol. Chem. 275 (2000) 18739-18744]. We have isolated genomic DNA clones covering the gene for the human DNA polymerase delta 50 kDa subunit (POLD2) and its 5'-flanking sequence. The POLD2 gene is composed of 11 exons and is distributed over 10 kb of genomic DNA. All exon-intron splice junctions conformed to the GT/AG consensus sequence. The 5'-flanking region of human POLD2 is G+C-rich and does not have a typical TATA box. A computer-based search for potential transcription factor binding sites revealed the existence of a number of motifs including those for AP1, AP2, Sp1, NF-1 and CREB. The functional activity of the regulatory region of the human POLD2 gene was demonstrated by its ability to drive the expression of a chloramphenicol acetyltransferase reporter gene in COS-7 cells.

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Sigma-Aldrich
Taq DNA polimerasi, with 10× PCR reaction buffer containing MgCl2
Sigma-Aldrich
Taq DNA polimerasi, with 10× PCR reaction buffer without MgCl2
Sigma-Aldrich
DNA Polymerase I from Escherichia coli lysogenic for NM 964, buffered aqueous glycerol solution