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[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.