Langmuir Monolayer as a Tool toward Visualization of a Specific DNA-Protein Complex

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• 作者：Loveleen K. Brar ; Priya Rajdev ; Arup K. Raychaudhuri ; and Dipankar Chatterji
• 刊名：Langmuir
• 出版年：2005
• 出版时间：November 8, 2005
• 年：2005
• 卷：21
• 期：23
• 页码：10671 - 10675
• 全文大小：671K
• 年卷期：v.21,no.23(November 8, 2005)
• ISSN：1520-5827

文摘

Immobilization and imaging of protein molecules and protein-DNA complexes on a Langmuir-Blodgett(LB) substrate have been explored here. We have prepared a nickel-arachidate (NiA) monolayer andcharacterized it through pressure-area isotherm on a LB trough. Recombinant RNA polymerase fromEscherichia coli, where the largest subunit was replaced with the same gene having a series of histidineamino acids at the C-terminus end of the protein, was immobilized over the Ni-arachidate monolayerthrough a Ni(II)-histidine interaction. A single molecule of RNA polymerase could be seen throughintermittent-contact atomic force microscopy (AFM). Under the condition of the formation of the LBmonolayer, the enzyme molecules were arrayed and transcriptionally active. Interestingly, they could pickup sequence specific DNA molecules from the subphase in an oriented fashion. On the other hand, preformedRNA polymerase Ni(II)-arachidate monolayers bound DNA haphazardly when no surface pressure wasemployed.