自稳定硫代磷酸寡核苷酸的合成及其在血液中的稳定性

Synthesis and stablization of self stablized phosphorothioate oligonucleotides in serum

目的：探讨自行设计的自稳定硫代磷酸寡核苷酸在血液中的稳定性。方法：在ＤＮＡ自动合成仪上合成硫代磷酸寡核苷酸和普通磷酸二酯键寡核苷酸，经高效液相仪纯化和３２Ｐ标记后，两者一起在２０％血清中３７℃温育０，１，４，８及３２ｈ，酚抽提后，变性聚丙烯酰胺凝胶电泳，比较两者的降解率。结果：自稳定硫代磷酸寡核苷酸在各时相点均为完整形式，普通磷酸二酯键寡核苷酸在以上５个时相点的降解率分别为０．０％，７７．２％，８３．０％，８４．４％和８５．３％。结论：自稳定硫代磷酸寡核苷酸在血液中有较强的稳定性。

Objective:Self stablized phosphorothioate oligonucleotides (PS oligos) were designed, and their stability in serum was detected.Methods: PS oligos and phosphodiester oligonucleotides (PDE oligos) were synthesized in automatic DNA syntheser, purified by high potential liquid chromatography,lablled by 32 P, and then incubated at 0,1,4,8 and 24 h respectively in 20% serum at 37 ℃.Extracted by phenol, deposition was electrophoresized in denatured polyacrylamide gel. Rates of mobility shift of PS oligos and PDE oligos in gel were assayed. Results: Degradation rates were all in PS oligos 0.0%,77.2%,83.0%,84.0% and 85.3% respectively in PDE oligos at above mentioned time points. Conclusion: Self stablized PS oligos have powerful resistance to nuclease in serum.