Synthetic promoter elements obtained by nucleotide sequence variation and selection for activity.

Synthetic promoter elements obtained by nucleotide sequence variation and selection for activity.
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通过核苷酸序列变异和活性选择获得的合成启动子元件。

DOI:
10.1073/pnas.97.7.3038
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发表时间:
2000
影响因子:
11.1
通讯作者:
F. S. Jones
F. S. Jones
中科院分区:
综合性期刊1区
文献类型:
--
作者:
G. Edelman;R. Meech;G. Owens;F. S. Jones

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不同细胞中的真核转录调控涉及大量顺式和反式元件及其排列。调查顺式调控元件的数量,在不同的情况下是活跃的,我们已经设计了一个高通量的选择程序,允许合成的活性顺式基序,提高了最小的启动子的活性。这种合成启动子构建方法(SPCM)用于鉴定在神经母细胞瘤细胞系Neuro 2A中显示出增加的启动子活性的>100个DNA序列。在确定所选合成启动子的DNA序列后,对已知元件的数据库搜索揭示了八个基序的优势:AP 2、CEBP、GRE、Ebox、ETS、CREB、AP 1和SP1/MAZ。所选合成启动子中最具活性的包含许多这些基序的复合物。使用三个示例性基序(ETS、CREB和SP1/MAZ)的DNA结合和启动子活性的测定来证明SPCM在揭示活性序列中的有效性。在目前可用的数据库中,133个选定的活性序列中有多达10%没有匹配,这提高了SPCM揭示新的基序和它们所结合的转录调控蛋白的可能性。该方法可用于构建活性顺式基序的数据库,用于诊断和基因治疗。
Eukaryotic transcriptional regulation in different cells involves large numbers and arrangements of cis and trans elements. To survey the number of cis regulatory elements that are active in different contexts, we have devised a high-throughput selection procedure permitting synthesis of active cis motifs that enhance the activity of a minimal promoter. This synthetic promoter construction method (SPCM) was used to identify >100 DNA sequences that showed increased promoter activity in the neuroblastoma cell line Neuro2A. After determining DNA sequences of selected synthetic promoters, database searches for known elements revealed a predominance of eight motifs: AP2, CEBP, GRE, Ebox, ETS, CREB, AP1, and SP1/MAZ. The most active of the selected synthetic promoters contain composites of a number of these motifs. Assays of DNA binding and promoter activity of three exemplary motifs (ETS, CREB, and SP1/MAZ) were used to prove the effectiveness of SPCM in uncovering active sequences. Up to 10% of 133 selected active sequences had no match in currently available databases, raising the possibility that new motifs and transcriptional regulatory proteins to which they bind may be revealed by SPCM. The method may find uses in constructing databases of active cis motifs, in diagnostics, and in gene therapy.
神经细胞粘附分子跨膜亚型的过度表达改变了大鼠 CNS-1 神经胶质瘤的侵袭性。
DOI: --
发表时间: 1998
期刊: Cancer research.
影响因子: --
作者:
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通讯作者: Kruse,CA
DOI: 10.1126/science.277.5334.2007
发表时间: 1997-09-26
期刊: SCIENCE
影响因子: 56.9
作者:
Valdivia, RH;Falkow, S
通讯作者: Falkow, S
DOI: 10.1126/science.8266075
发表时间: 1993-12-17
期刊: SCIENCE
影响因子: 56.9
作者:
LI, JJ;HERSKOWITZ, I
通讯作者: HERSKOWITZ, I
DOI: 10.1101/gad.12.1.34
发表时间: 1998-01-01
影响因子: 10.5
作者:
Lagrange, T;Kapanidis, AN;Ebright, RH
通讯作者: Ebright, RH
DOI: 10.1073/pnas.92.6.1955
发表时间: 1995-03-14
影响因子: 11.1
作者:
COLGAN, J;MANLEY, JL
通讯作者: MANLEY, JL