申请人:The Perkin-Elmer Corporation
公开号:US06008379A1
公开(公告)日:1999-12-28
A class of aromatic-substituted xanthene compounds useful as fluorescent dyes is disclosed, the compounds having the general structure where Y.sub.1 and Y.sub.2 taken separately are selected from the group consisting of hydroxyl, oxygen, imminium, linking group and amine, or Y.sub.1 taken together with R.sub.2 is cyclic imine, or Y.sub.2 taken together with R.sub.3 is cyclic amine; R.sub.2, R.sub.3, R.sub.5, and R.sub.7 taken separately are selected from the group consisting of hydrogen, fluorine, chlorine, lower alkyl, lower alkene, lower alkyne, sulfonate, sulfone, amino, imminium, amido, nitrile, lower alkoxy, phenyl, and linking group; R.sub.1 taken separately is selected from the group consisting of phenyl, substituted phenyl, polycyclic aromatic, substituted polycyclic aromatic, linking group and electron-rich heterocycle, or when taken together with R.sub.7 is selected from the group consisting of electron-rich heterocycle and indene; R.sub.4 taken separately is selected from the group consisting of amino, amido, phenyl, substituted phenyl, polycyclic aromatic, substituted polycyclic aromatic, indene, linking group and electron-rich heterocycle, or when taken together with R.sub.5 is selected from the group consisting of phenyl, substituted phenyl, polycyclic aromatic, substituted polycyclic aromatic, indene, and electron-rich heterocycle; and R.sub.6 is selected from the group consisting of acetylene, lower alkyl, lower alkene, cyano, phenyl, substituted phenyl, and heterocyclic aromatic. In another aspect, the invention includes methods for synthesizing the above dye compounds and intermediates. In yet another aspect, the present invention includes reagents labeled with the asymmetric benzoxanthene dye compounds, including deoxynucleotides, dideoxynucleotides, phosphoramidites, and polynucleotides. In an additional aspect, the invention includes methods utilizing such dye compounds and reagents including dideoxy polynucleotide sequencing and fragment analysis methods.
揭示了一类芳香基取代的黄酮类化合物,可用作荧光染料,其中化合物具有一般结构式,其中Y1和Y2分别选自羟基、氧、亚胺、连接基和胺的群组,或Y1与R2一起选为环状亚胺,或Y2与R3一起选为环状胺;R2、R3、R5和R7分别选自氢、氟、氯、低碳基、低碳烯、低碳炔、磺酸酯、磺酮、氨基、亚胺、酰胺、腈、低碳氧基、苯基和连接基;R1分别选自苯基、取代苯基、多环芳香族、取代多环芳香族、连接基和电子富集杂环,或与R7一起选自电子富集杂环和茚;R4分别选自氨基、酰胺、苯基、取代苯基、多环芳香族、取代多环芳香族、茚、连接基和电子富集杂环,或与R5一起选自苯基、取代苯基、多环芳香族、取代多环芳香族、茚和电子富集杂环;R6选自乙炔、低碳基、低碳烯、氰基、苯基、取代苯基和杂环芳香族。在另一个方面,本发明包括合成上述染料化合物和中间体的方法。在另一个方面,本发明包括用不对称苯并黄酮染料化合物标记的试剂,包括脱氧核苷酸、二脱氧核苷酸、磷酰胺酰胺和多核苷酸。在另一个方面,本发明包括利用这种染料化合物和试剂的方法,包括二脱氧多核苷酸测序和片段分析方法。