[Study on cis-acting elements of rat glutathione S-tranferase P1 gene].
OBJECTIVE
To investigate the cis-acting regulatory DNA elements in 3.0 kb 5' flanking upstream region of rat glutathione S-tranferase (rGSTP1) gene.


METHODS
Using exonuclease III (Exo III) and S1 nuclease, a series of deletions were made in the 3.0 kb region, then fused into highly sensitive luciferase reporter vector. The deleted mutants were transfected into HeLa cells. Luciferase activities of the transfected cell lysates were assayed.


RESULTS
Deleted from -2.9 kb to -2.3 kb, luciferase activity had dropped about 67% (P < 0.05). The element between -2.5 kb and -2.2 kb could increase gene expression activity regardless of orientation and position. Further deleted from -1.8 kb to -1.3 kb, luciferase activity increased about three times (P < 0.05).


CONCLUSIONS
There are enhancer element between -2.5 kb and -2.2 kb and a negative regulatory sequence between -1.8 kb and -1.3 kb in the upstream region of rGSTP1 gene.
Liu
D
D
Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005.
Quan
S
S
Wang
J
J
Fang
F
F
D000818:Animals; D004742:Enhancer Elements, Genetic; D017930:Genes, Reporter; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D008156:Luciferases / Q000235:genetics; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D014162:Transfection
D007527:Isoenzymes; D008156:Luciferases; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase; C496558:Gstp1 protein, rat