| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
Terminal modification for oligonucleotides is necessary for immobilization of the oligonucleotides on solid surface. There are many kinds of terminal modification such as sulfhydryl group, hydrazide, anthraquinone and acrylamide ; among them, the aliphatic amine is most cost effective and produces less side products under standard deprotecting condition after the oligonucleotide synthesis. Thus, the oligonucleotides modified with aliphatic amine have been frequently used for a gene analysis system requiring diverse oligonucleotide probes. Besides the aliphatic amino groups, there are nucleosidic type amino-modifiers to date. However, these nucleosidic amino-modifiers are able to form base pairs with opposite nucleotide in binding with complementary strand. Therefore, these nucleosidic amino-modifiers are not suitable as universal modification for diverse oligonucleotide probes.
Then, we developed novel amino-modifiers, which increased the reactivity and improved purification process of the amino-modified oligonucleotides. We designed and synthesized a series of amidite units, which have a primary amino group and an aromatic residue by connecting with a short or a long linker of various lengths. Naphthalene and anthracene were selected as the aromatic groups. The amino-modifiers containing the aromatic residues showed higher reactivity than the conventional amino-modification. Especially, the amino-modifier of which the primary amine was proximal to the hydrophobic residue had a high reactivity. Furthermore, these amino-modified oligonucleotides efficiently ligated to oligonucleotides whose 3'-end was oxidized by periodate ions. This crosslinking reaction proceeded on surface of solid phase.
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