C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01) C07K 14/775 (2006.01) C12N 5/10 (2006.01) C12N 15/11 (2006.01)
Patent
CA 2159532
Methods for screening for a drug that increases expression of the human apolioprotein (apo) Al gene and related DNA constructs. A first method has the steps of: (a) introducing into mammalian cells a DNA construct including, functionally joined together in the 5'- 3' direction of transcription, (i) at least one copy of a drug-responsive element (DRE) having at least about 80% homology with the DNA sequence 5'-G/C N T/G A/G GCTGGG-3', (ii) a heterologous promoter, (iii)a reporter gene and (iv) an untranslated region including a functional polyadenylation signal; (b) growing a first culture of the cells in the absence of drug; (c) lysing the first culture to produce a first extract; (d) assaying the first extract for activity of a protein encoded by the reporter gene; (e) growing a second culture of the cells in the presence of the drug; (f) lysing the second culture to produce a second extract; (g) assaying the second extract for activity of the protein encoded by the reporter gene; and (h) comparing the activities of the first extract and the second extract. A second method uses a different DNA construct including (i) a promoterregion of the human apo Al gene including at least one copy of a drug-responsiveelement (DRE) having at least about 80% homology with the DNA sequence 5'-G/C N T/G A/G GCTGGG-3', (ii) a reporter gene and (iii) an untranslated region including a functional polyadenylation signal. A third method involves stably maintaining the introduced DNA in the cells.
Tam Shui-Pang
Zhang Xia
Queen's University At Kingston
Steeg Carol Miernicki
LandOfFree
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