Sensitivity of virally-driven luciferase reporter plasmids to members of the steroid/thyroid/retinoid family of nuclear receptors
Sensitivity of virally-driven luciferase reporter plasmids to members of the steroid/thyroid/retinoid family of nuclear receptors
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DOI：
10.1016/s0960-0760(99)00109-0
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发表时间：
1999-09-01
影响因子：
4.1
通讯作者：
Crabb, DW
中科院分区：
文献类型：
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作者：
Everett, LM;Crabb, DW
在一系列转染实验中，发现用作内部对照的pRSV-luc质粒对与类固醇/甲状腺/类维生素A核受体超家族的几个成员的表达载体的共转染敏感。因此，进行了这些表达载体对四种报告质粒活性的影响的调查。在CV-1细胞中，含有P. pyralis荧光素酶基因的pRSV-luc的活性被共转染的PPAR α和阿普-1抑制，并被COUP-TFI激活。含有相同荧光素酶基因的pSV 40-luc的表达被PPARa和HNF-4抑制，并被COUP-TFI和阿普-1激活。所有这四种表达载体都降低了pRL-TK质粒的表达，该质粒含有来自海肾的荧光素酶基因。RXR表达载体在CV-I细胞中对荧光素酶活性没有影响，但在H4 IIEC 3肝癌细胞中诱导荧光素酶活性。这种激活被阻断的配体，9-顺式视黄酸的加入。含有氯霉素乙酰转移酶基因的pSV 2-CAT对所有测试的受体表达载体均不敏感。P. pyralis和R.肾形萤光素酶基因似乎含有使它们对类固醇/甲状腺/类维生素A核受体有反应的序列。(C)1999 Elsevier Science Ltd.保留所有权利。
During a series of transfection experiments, the pRSV-luc plasmid used as an internal control was found to be sensitive to co-transfection with expression vectors for several members of the steroid/thyroid/retinoid superfamily of nuclear receptors, Therefore, a survey of the effect of these expression vectors on the activity of four reporter plasmids was conducted. In CV-1 cells, the activity of pRSV-luc, which contains the P. pyralis luciferase gene, was repressed by co-transfection of PPAR alpha and ARP-1 and was activated by COUP-TFI. Expression of pSV40-luc, containing the same luciferase gene, was repressed by PPARa and HNF-4 and activated by both COUP-TFI and ARP-1. All four of these expression vectors reduced the expression of the pRL-TK plasmid, which contains the luciferase gene from Renilla reniformis. RXR expression vectors had no effect on luciferase activity in CV-I cells but induced luciferase activity in H4IIEC3 hepatoma cells. This activation was blocked by the addition of ligand, 9-cis retinoic acid. pSV2-CAT, which contains the chloramphenicol acetyltransferase gene, was insensitive to all receptor expression vectors tested. Both the P. pyralis and R. reniformis luciferase genes appear to contain sequences that render them responsive to steroid/thyroid/retinoid nuclear receptors. (C) 1999 Elsevier Science Ltd. All rights reserved.