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首页> 外文期刊>Current Protocols in Nucleic Acid Chemistry >Synthesis of 2'-Deoxyuridine Modified with a 3,5-Difluoro-4-Methoxybenzylidene Imidazolinone Derivative for Incorporation into Oligonucleotide Probes for Detection of HER2 Breast Cancer Marker
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Synthesis of 2'-Deoxyuridine Modified with a 3,5-Difluoro-4-Methoxybenzylidene Imidazolinone Derivative for Incorporation into Oligonucleotide Probes for Detection of HER2 Breast Cancer Marker

机译：2脱氧尿苷的合成与修改3, 5-Difluoro-4-Methoxybenzylidene Imidazolinone导数并入寡核苷酸探针检测HER2乳腺癌标记

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Nucleoside intercalator conjugates (NICs) describe an innovative methodology developed in our research group for preparation of fluorescence turn-on DNA hybridization probes targeting specific mRNA sequences (e.g., breast cancer markers). In this methodology, we conjugate a non-fluorescent inter-calator to the base of a nucleic acid (e.g., uracil) via a flexible spacer. This modified monomer can be incorporated into oligonucleotides by solid-phase synthesis and a large fluorescence enhancement is observed when the modified oligonucleotide is hybridized with its complementary strand due to intercalation of the fluorophore between the two strands. 5-(6-p-Methoxybenzylidene imidazolinone-1-hexene)-2;-deoxyuridine (dUMBI) is a synthetic monomer to which4-methoxybenzylidene imidazolinone (MBI), the fluorescent chro-mophore of green fluorescent protein (GFP), has been conjugated via a flexible spacer. The detection of human epidermal growth factor receptor 2 (HER2) mRNA by this probe has already been established by our group. The fluorescent intensity of the single-strand DNA can be considered as negligible due to the free rotation of the fluorophore. Upon hybridization, however, the flexible spacer allows for the intercalation of the fluorophore between the hybridized strands, giving rise to enhanced fluorescence and indicating the presence of target mRNA. 3,5-Difluoro-4-methoxybenzylidene (DFMBI) has enhanced photophysical properties compared to MBI fluorophore. This protocol describes a simple, reliable, efficient, and general method for the synthesis of improved derivative dUDFMBI as a monomer of fluorescent turn-on DNA hybridization probe with application for detection of HER2 mRNA.
机译：核苷intercalator轭合物(nic)描述在我们开发的一种创新方法研究小组准备的荧光刺激DNA杂交探针定位特定的信使rna序列(例如,乳腺癌标记)。荧光inter-calator的基地核酸(例如,尿嘧啶)通过一个灵活的垫片。固相合成的寡核苷酸和一个大观察到荧光增强当修改后的寡核苷酸杂交由于其互补链两者之间的荧光团的夹层链。imidazolinone-1-hexene) 2;脱氧尿苷(dUMBI)是合成单体which4-methoxybenzylidene imidazolinone (MBI)绿色荧光的荧光chro-mophore蛋白(GFP),通过一个灵活的共轭垫片。因子受体2 (HER2) mRNA探测器已经建立了我们的团队。单链DNA的荧光强度可以被认为是可以忽略不计由于自由荧光团的旋转。然而,柔性间隔允许夹层之间的荧光团杂化链,增强荧光和指示的存在目标mRNA。(DFMBI)增强的光物理性质而MBI荧光团。描述了一个简单、可靠、高效和一般的合成方法改进导数dUDFMBI单体的荧光刺激DNA杂交探针与应用程序检测HER2 mRNA。

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