scholarly journals A DNA-binding protein factor in K562 nuclear extract interacts with positive control region (PCR) in the 5′-flanking sequence of human β-globin gene

Cell Research ◽  
1993 ◽  
Vol 3 (1) ◽  
pp. 103-110
Author(s):  
Yulong Hu ◽  
Yadi Chen ◽  
Tong Sun ◽  
Ruolan Qian
Keyword(s):  
Dna Binding ◽  
Control Region ◽  
Binding Protein ◽  
Globin Gene ◽  
Dna Binding Protein ◽  
Nuclear Extract ◽  
Positive Control ◽  
Flanking Sequence ◽  
Protein Factor

Diurnal regulation of a DNA binding protein to the period repeat sequence in the SCN nuclear extract of rat brain

1999 ◽  
Vol 65 (2) ◽  
pp. 211-215 ◽  
Author(s):  
Toshiyuki Hamada ◽  
Koichiro Kako ◽  
Hisanori Wakamatsu ◽  
Shigenobu Shibata ◽  
Shigenori Watanabe ◽  
...  
Keyword(s):  
Dna Binding ◽  
Rat Brain ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Nuclear Extract ◽  
Repeat Sequence ◽  
Diurnal Regulation

scholarly journals The rat H+/K(+)-ATPase beta subunit gene and recognition of its control region by gastric DNA binding protein.

1991 ◽  
Vol 266 (32) ◽  
pp. 21584-21588 ◽  
Author(s):  
M. Maeda ◽  
K. Oshiman ◽  
S. Tamura ◽  
S. Kaya ◽  
S. Mahmood ◽  
...  
Keyword(s):  
Dna Binding ◽  
Control Region ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Beta Subunit ◽  
Subunit Gene ◽  
Beta Subunit Gene

scholarly journals SpZ12-1, a negative regulator required for spatial control of the territory-specific CyIIIa gene in the sea urchin embryo

Development ◽  
1995 ◽  
Vol 121 (4) ◽  
pp. 1111-1122 ◽  
Author(s):  
D.G. Wang ◽  
C.V. Kirchhamer ◽  
R.J. Britten ◽  
E.H. Davidson
Keyword(s):  
Dna Binding ◽  
Sea Urchin ◽  
Fusion Gene ◽  
Binding Protein ◽  
Regulatory Element ◽  
Regulatory Region ◽  
Dna Binding Protein ◽  
Nuclear Extract ◽  
Negative Regulator ◽  
Expression Library

The CyIIIa cytoskeletal actin gene of the sea urchin Strongylocentrotus purpuratus is activated in late cleavage and expressed exclusively in the aboral ectoderm territory of the embryo. Previous gene transfer studies defined a 2.3 kb cis-regulatory region that is necessary and sufficient for correct temporal and spatial expression of a CyIIIa.CAT fusion gene. In this paper, a negative regulatory element within this region was identified that is required for repression of the CyIIIa gene in skeletogenic mesenchyme cells. The repression mediated by this regulatory element takes place after initial territorial specification. A cDNA clone encoding a DNA-binding protein with twelve Zn fingers (SpZ12-1) was isolated by probing an expression library with this cis-element. Deletion analysis of the SpZ12-1 protein confirmed that a DNA-binding domain is located within the Zn finger region. SpZ12-1 is the only DNA-binding protein in embryo nuclear extract that interacts with the specific cis-target sites required for repression of CyIIIa.CAT in skeletogenic mesenchyme and is likely to be the trans factor that mediates this repression.

scholarly journals Development of DNA Pair Biosensor for Quantization of Nuclear Factor Kappa B

Biosensors ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 126
Author(s):  
Zhaohui Wang ◽  
Pak Wong
Keyword(s):  
Dna Binding ◽  
Nuclear Factor Kappa B ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Nuclear Extract ◽  
Molecular Probe ◽  
Nuclear Factor ◽  
Hela Nuclear Extract ◽  
Molecular Binding ◽  
Nuclear Factor Kappa

Nuclear factor kappa B (NF-κB), regulating the expression of several genes that mediate the inflammatory responses and cell proliferation, is one of the therapeutic targets for chronic inflammatory disease and cancer. A novel molecular binding scheme for the detection of NF-κB was investigated for its affinity to Ig-κB DNA composed by dye and quencher fluorophores, and this specificity is confirmed by competing with the DNA sequence that is complementary to the Ig-κB DNA. We create a normalization equation to remove the negative effects from the various initial fluorophore concentrations and the background noise. We also found that a periodic shaking at a frequency could help to stabilize the DNA–protein binding. The calibration experiment, using purified p50 (NF-κB), shows that this molecular probe biosensor has a detection limit on the order of nanomolar. The limit of detection is determined by the binding performance of dye and quencher oligonucleotides, and only a small portion of probes are stabilized by DNA-binding protein NF-κB. The specificity experiment also shows that p50/p65 heterodimer has the highest affinity for Ig-κB DNA; p65 homodimer binds with intermediate affinity, whereas p50 shows the lowest binding affinity, and Ig-κB DNA is not sensitive to BSA (bovine albumin serum). The experiment of HeLa nuclear extract shows that TNF-α stimulated HeLa nuclear extract has higher affinity to Ig-κB DNA than non-TNF-stimulated HeLa nuclear extract (4-h serum response). Therefore, the molecular binding scheme provides a rapid, quantitative, high throughput, and automated measurement of the DNA-binding protein NF-κB at low cost, which is beneficial for automated drug screening systems.

scholarly journals In Vitro Methylation of theBsuRI (5′-GGCC-3′) Sites in the E2a Region of Adenovirus Type 2 DNA Does Not Affect Expression inXenopus laevisOocytes

1982 ◽  
Vol 2 (12) ◽  
pp. 1574-1580 ◽  
Author(s):  
Lily Vardimon ◽  
Ursula Günthert ◽  
Walter Doerfler
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
Dna Binding ◽  
Cell Lines ◽  
Control Region ◽  
Binding Protein ◽  
Adenovirus Type ◽  
Dna Binding Protein

The early region 2a (E2a) of adenovirus type 2 (Ad2) DNA codes for a 72,000-dalton DNA-binding protein and is expressed in the Ad2-transformed hamster cell line HE1 but not in cell lines HE2 and HE3 (H. Esche, J. Virol.41:1076-1082, 1982; K. Johansson et al., J. Virol.27:628-639, 1978). An inverse correlation between DNA methylation at the 5′-CCGG-3′ sites of the E2a region and of gene expression in these cell lines has been observed (L. Vardimon et al., Nucleic Acids Res.8:2461-2473, 1980). When the cloned E2a region of Ad2 DNA is methylated in vitro at the 5′-CCGG-3′ sites, the gene is not transcribed after being injected into the nuclei ofXenopus laevisoocytes, whereas unmethylated DNA is expressed (L. Vardimon et al., Eur. J. Cell Biol.25:13-15, 1981; L. Vardimon et al., Proc. Natl. Acad. Sci. U.S.A.79:1073-1077, 1982). These data demonstrate that DNA methylation is directly involved in the shut-off of transcription. In the present communication we investigated in detail the control region of the gene for the DNA-binding protein in Ad2-transformed cell lines and showed that the first late control region (map coordinate 72 on the viral DNA) of the E2a region is present in its entirety in cell lines HE1, HE2, and HE3. TheHaeIII sites (5′-GGCC-3′) in the E2a region in all three cell lines were not methylated. When the DNA methyltransferaseBsuRI was used, all 5′-GGCC-3′ sites in the cloned E2a region of Ad2 DNA were methylated in vitro. It was shown that methylation of these sites did not inhibit the expression of this viral gene inX. laevisoocytes. Thus, for methylation to affect gene expression in the E2a region it has to occur at specific sites (e.g., 5′-CCGG-3′) which may be different for other genes.

Sign in / Sign up

Export Citation Format

Share Document