Nagashima Jun-Ichi, Minezaki Satoshi, Obika Satoshi, Imanishi Takeshi, Kuwahara Masayasu, Sawai Hiroaki
Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515, Japan.
Nucleic Acids Symp Ser (Oxf). 2007(51):55-6. doi: 10.1093/nass/nrm028.
We have attempted to synthesise a complimentary strand using oligo-DNA with modified bases, sugars and phosphates as a template strand by polymerase reaction. Analogues bearing C5-substituted uracil, those with amide linkage [-CH2C=ONH-] in place of phospho-diester linkage and those bearing 2'-O, 4'-C-bridged sugar were used. Primer extension reactions were carried out to synthesise complimentary DNA strands. The reactions depended on the thermostable DNA polymerase used, the type of modification or the number of the modified position on the template strand.
我们尝试通过聚合酶反应，以带有修饰碱基、糖和磷酸的寡聚DNA作为模板链来合成互补链。使用了带有C5取代尿嘧啶的类似物、用酰胺键[-CH2C=ONH-]取代磷酸二酯键的类似物以及带有2'-O, 4'-C-桥连糖的类似物。进行引物延伸反应以合成互补DNA链。这些反应取决于所使用的热稳定DNA聚合酶、修饰的类型或模板链上修饰位置的数量。
