固體支持亞磷酸三脂法合成寡去氧核醣核苷酸d-AATT與d-GGCC

Chemical Synthesis of d-AATT and d-GGCC by Phosphite Triester Method

許宗雄(T. H. Hseu) ； 甘魯生(L. S. Kan)

《化學》 42卷2期 (1984/06) Pp. 77-82

https://doi.org/10.6623/chem.1984011

摘要

本研究利用固體支持亞磷酸三脂法合成二段去氧核醣核苷酸d-AATT與d-GGCC。終產物經高速液相層析分離純化，繼利用蛇毒磷酸雙脂酶分解鑑定，顯示反應進行良好。d-AATT之声率約爲75%，卽每一合成步驟效率高於90%。而d-GGCC產率較低，約爲45 %，由於副物極少，d-GGCC低產率可歸因於dC之固體支持物於運輸與保存情況下有部份分解之故。雖如此，由本試驗可知在國內潮濕氣候下於有溫調的普通化學實驗室可成功的進行寡去氧核醣核苷酸的化學合成。有關合成步驟於文中有詳細描述。

並列摘要

The synthesis of short deoxyribonucleotides (DNA) with de sired sequences and high purity is of prime importance to the rapid developments of molecular biology which is a major branch of the life sciences. Both, phosphotriester and phosphite triester methods can provide an efficient way to produce short DNA (RNA as well. However, the experience tells us both reactions will run well under extreme anhydrous condition. In order to find how well these condensations can be performed in a very humid environment such as Taiwan, a subtropical island, two tetramer, d-AATT-and d-GGCC ,were synthesized in an ordinary chemical hood of an air-conditioned laboratory, The result of high pressure liquid chromatograph has shown that reaction went completed in both trials There is no significant impurities found and the efficiency of each nucleotidyl addition step is calculated as 90%+ for d-AAT which is acceptable, and only 75% for d-GGCC. The low efficiency of each addition in the case of d-GGCC may have resulted from the deterioration of the starting materials, especially the solid-bound protected nucleosides, by discovering yellowish staff in solid support deoxycytidine Nonetheless this pilot trial of short DNA synthesis can be considered as successful A detailed step-by-step description of the methods is given.