pE2F-TA-Luc

pE2F-TA-Luc

pE2F-TA-Luc载体基本信息：

pE2F-TA-Luc载体质粒图谱和多克隆位点信息：

pE2F-TA-Luc载体简介：

pE2F-TA-Luc is designed to monitor the induction of E2F-mediated signal transduction pathways.

E2F, a major target of the retinoblastoma gene (Rb), is a key regulator of cell-cycle checkpoints

in mammalian cells (1–2). E2F plays a critical role in stimulating expression of genes encoding

growth-promoting proteins (3), and is involved in regulating the expression of important genes

during cell proliferation (4–5). The E2F protein forms a heterodimer complex with the DP1

protein, which binds to E2F response elements and initiates transcription of genes necessary

for DNA replication. Studies have shown that deregulation of E2F results in a loss of cell-cycle

checkpoints—thereby predisposing cells to uncontrolled growth (1–5). pE2F-TA-Luc contains

four copies of the E2F enhancer element (6), located upstream of the minimal TA promoter, the

TATA box from the herpes simplex virus thymidine kinase promoter (PTA). Located downstream

of PTA is the firefly luciferase reporter gene (luc). Upon binding of the E2F/DP1 complex to the

cis-acting E2F enhancer element, transcription is induced and the reporter gene is activated.

The luciferase coding sequence is followed by the SV40 late polyadenylation signal to ensure

proper, efficient processing of the luciferase transcript in eukaryotic cells. A synthetic transcription

blocker (TB) is located upstream of the cis-acting enhancer element. It is composed of

adjacent polyadenylation and transcription pause sites for blocking nonspecific transcription

(7). The vector backbone also contains an f1 origin for single-stranded DNA production, a pUC

origin of replication, and an ampicillin resistance gene for propagation and selection in E. coli.

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