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Several ligases and firefly luciferase catalyze the synthesis of dinucleoside polyphosphates

100%

Sillero A., Sillero M. A. G.

Cellular and Molecular Biology Letters

1999

tom 04

nr 3

Control of phenylalanine ammonia-lyase gene promoters from pea by UV radiation

86%

Pluskota W. E., Michalczyk D. J., Gorecki R. J.

Acta Physiologiae Plantarum

2005

tom 27

nr 2

The gene fusion system was used to study UV light-control of PS PAL1 and PS PAL2 genes encoding phenylalanine ammonia-lyase of pea. The induction of pea PAL promoters was analysed in transgenic tobacco plants. Binary plasmids (derivatives of pBI101.2 vector) containing 5’ regulatory fragments of PS pAl1 and PS PAL2 linked to reporter genes (GUS, LUC) were constructed. The analyses were performed with the use of single constructs (containing one variant of PS PAL promoter and one reporter gene) and dual constructs (containing both PS PAL1 and PS PAL2 promoters connected with different reporter genes). The use of dual constructs enabled the evaluation of both PS PAL promoters activity in the same plant. The analyses of in vitro grown plants have shown that both PAL promoters are strongly induced in leaves subjected to UV radiation. In some cases, the UV-stimulated expression exceeded the exposed areas. This phenomenon was observed more often in the leaves of plants containing the PS PAL1:: GUS than PS PAL2::GUS construct. Removal ofboxes 2, 4, 5 from PS PAL1 promoter and deletion of its 5’end region (-339 to -1394) decreases the level of gene expression but does not eliminate its responsiveness to UV.

Molecular cloning and analysis of the human PCAN1 [GDEP] promoter

72%

Liu W., Chen W., Zhang P., Yu C., Kong F., Deng J., Zhang J., Jiang A.

Cellular and Molecular Biology Letters

2007

tom 12

nr 4

482-492

Human PCAN1 (prostate cancer gene 1) is a prostate-specific gene that is highly expressed in prostate epithelial tissue, and frequently mutated in prostate tumors. To better understand the regulation of the PCAN1 gene, a 2.6-kb fragment of its 5′ flanking region was obtained by PCR. Its promoter activity was examined via the dual-luciferase reporter assay after it had been cloned into a pGL3-basic vector generating pGL3-p2.6kb and transfected into LNCaP cells. pGL3-basic and pGL3-control were respectively used as the negative and positive controls. Sequence analysis with the MatInspector database showed that some possible binding sites for the transcriptional factors, NKX3.1, P53, SP1, cEBP and the PPAR/RXR heterodimers may locate on a 2.6-kb region upstream of the PCAN1 gene. To examine the relevant regulation of PCAN1, pGL3-p2.6kb was transfected into the prostate cancer cell line LNCaP, which was treated with R1881 (10−7∼10−9 mol/l), 17β-estradiol (17β-E2, 10−7∼10−9 mol/l), all-trans-retinoic acid (all-trans-RA, 10−5∼10−7 mol/l) or 9-cis-retinoic acid (9-cis-RA, 10−5∼10−7 mol/l), and eukaryotic expression plasmids of NKX3.1, p53, Sp1, Pten, PPARγ or cEBPα were cotransfected with pGL3-p2.6kb into LNCaP cells. pRL-TK, a Renilla luciferase reporter vector, was cotransfected into all the transfection lines as an internal control. The activities of pGL3-p2.6kb (PCAN1 promoter) were analyzed via the dual-luciferase reporter assay 48 h after transfection. The results showed that 9-cis-RA enhanced the PCAN1 promoter activity in a dose-dependent manner, while R1881, 17β-E2 and all-trans-RA had no significant effect on PCAN1 promoter activities. Cotransfection with pGL3-p2.6kb and the expression plasmids of NKX3.1, p53, Sp1 or Pten respectively resulted in 1.66-, 2.48-, 2.00-and 1.72-fold 2.6 kb PCAN1 promoter activity increases relative to the controls, which were cotransfected with pcDNA3.1(+), while cotransfection of PPARγ and cEBPα yielded no significant effect on PCAN1 promoter activities. These results could be applied for further study of the function and transcription regulation of the PCAN1 gene in prostate development and carcinogenesis.

Ograniczanie wyników

2 Cellular and Molecular Biology Letters

1 Acta Physiologiae Plantarum

1 Chen W.

1 Deng J.

1 Gorecki R. J.

1 Jiang A.

1 Kong F.

1 Liu W.

1 Michalczyk D. J.

1 Pluskota W. E.

1 Sillero A.

1 Sillero M. A. G.

1 Yu C.

1 Zhang J.

1 Zhang P.

1 2007

1 2005

1 1999

Wyniki wyszukiwania

Several ligases and firefly luciferase catalyze the synthesis of dinucleoside polyphosphates

Control of phenylalanine ammonia-lyase gene promoters from pea by UV radiation

Molecular cloning and analysis of the human PCAN1 [GDEP] promoter