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Volumn 130, Issue 10, 2008, Pages 2924-2925

Electrical detection of TATA binding protein at DNA-modified microelectrodes

Author keywords

[No Author keywords available]

Indexed keywords

DNA; TATA BINDING PROTEIN;

EID: 41449104768     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja7106756     Document Type: Article
Times cited : (83)

References (47)
  • 32
    • 41449089056 scopus 로고    scopus 로고
    • DNA was synthesized using standard phosphoramidite chemistry and characterized by MALDI-TOF mass spectrometry, UV-visible spectroscopy, and HPLC The 5′c C6 Thiol Modifier S-S and NHS-Carboxy-dT phosphoramidites were purchased from Glen Research. On solid support, NB was coupled to the 5-[3-acrylate NHS Ester]-deoxy uridine to form an amide bond. See Supporting Information for the NB coupling and characterization
    • DNA was synthesized using standard phosphoramidite chemistry and characterized by MALDI-TOF mass spectrometry, UV-visible spectroscopy, and HPLC The 5′c C6 Thiol Modifier S-S and NHS-Carboxy-dT phosphoramidites were purchased from Glen Research. On solid support, NB was coupled to the 5-[3-acrylate NHS Ester]-deoxy uridine to form an amide bond. See Supporting Information for the NB coupling and characterization.
  • 33
    • 0035171821 scopus 로고    scopus 로고
    • Significant hypochromicity of the NB chromophore is seen in the NB-DNA conjugate which likely indicates that NB is stacked in the duplex Supporting Information, For further spectroscopic characterization of the interaction of free NB with DNA, see: Lakowicz, J. R, Piszczek, G, Kang, J. S. Anal. Biochem. 2001, 288, 62
    • Significant hypochromicity of the NB chromophore is seen in the NB-DNA conjugate which likely indicates that NB is stacked in the duplex (Supporting Information). For further spectroscopic characterization of the interaction of free NB with DNA, see: Lakowicz, J. R.; Piszczek, G.; Kang, J. S. Anal. Biochem. 2001, 288, 62.
  • 35
    • 41449113151 scopus 로고    scopus 로고
    • We have found that the midpoint potential of NB-DNA monolayers on macroelectrodes vary depending upon the gold surface utilized, leading to an uncertainty of ∼50 mV in the potentials, although they have a higher certainty within a given surface. Moreover, there is a systematic negative shift in potential of ∼100 mV on the microelectrodes versus the macroelectrodes. Comparable shifts have been seen by others (see ref 18 b,c) and may reflect the different geometries of DNA monolayers on the different surfaces. See Supporting Information for further details
    • We have found that the midpoint potential of NB-DNA monolayers on macroelectrodes vary depending upon the gold surface utilized, leading to an uncertainty of ∼50 mV in the potentials, although they have a higher certainty within a given surface. Moreover, there is a systematic negative shift in potential of ∼100 mV on the microelectrodes versus the macroelectrodes. Comparable shifts have been seen by others (see ref 18 b,c) and may reflect the different geometries of DNA monolayers on the different surfaces. See Supporting Information for further details.
  • 37
    • 85153002467 scopus 로고    scopus 로고
    • At small scales, we observe a partially sigmoidal shape for the voltammetry of Nile Blue-DNA films. We attribute the sigmoidal shape to a small catalytic contribution to the NB signal from trace oxygen, as reported in Ju, H, Shen, C. Electroanalysis 2001, 13, 789. See Supporting Information
    • At small scales, we observe a partially sigmoidal shape for the voltammetry of Nile Blue-DNA films. We attribute the sigmoidal shape to a small catalytic contribution to the NB signal from trace oxygen, as reported in Ju, H.; Shen, C. Electroanalysis 2001, 13, 789. See Supporting Information.
  • 38
    • 41449110853 scopus 로고    scopus 로고
    • See Supporting Information for details of protein preparation.
    • See Supporting Information for details of protein preparation.
  • 41
    • 85177017971 scopus 로고    scopus 로고
    • At low TBP concentrations, where TBP binds site specifically, we observe little attenuation on electrodes modified with NB-DNA lacking the binding site. For binding affinities, see: (a) Coleman, R. A, Taggart, A. K. P, Benjamin, L. R, Pugh, B. F. J. Biol. Chem. 1995, 270, 13842
    • At low TBP concentrations, where TBP binds site specifically, we observe little attenuation on electrodes modified with NB-DNA lacking the binding site. For binding affinities, see: (a) Coleman, R. A.; Taggart, A. K. P.; Benjamin, L. R.; Pugh, B. F. J. Biol. Chem. 1995, 270, 13842.
  • 43
    • 0035353388 scopus 로고    scopus 로고
    • BSA is known to adsorb nonspecifically on self-assembled alkanethiol monolayers. For example, see:, Here we utilize BSA effectively to passivate the Au surface against further nonspecific adsorption
    • BSA is known to adsorb nonspecifically on self-assembled alkanethiol monolayers. For example, see: Holmlin, R. E.; Chen, X.; Chapman, R. G.; Takayama, S.; Whitesides, G. M. Langmuir 2001, 17, 2841. Here we utilize BSA effectively to passivate the Au surface against further nonspecific adsorption.
    • (2001) Langmuir , vol.17 , pp. 2841
    • Holmlin, R.E.1    Chen, X.2    Chapman, R.G.3    Takayama, S.4    Whitesides, G.M.5


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.