Taylor J S, Brockie I R
Department of Chemistry, Washington University, St Louis, MO 63130.
Nucleic Acids Res. 1988 Jun 10;16(11):5123-36. doi: 10.1093/nar/16.11.5123.
The synthesis of a building block for the sequence specific introduction of the trans-syn thymine dimer into oligonucleotides via solid phase DNA synthesis technology is described. CGTAT[t,s]TATGC was synthesized in 48% overall yield by a partially automated procedure. The stepwise coupling yield for addition of the trans-syn thymine dimer building block was 58%. The dimer containing oligonucleotide was characterized by 500 MHz 1H COSY and NOESY spectroscopy and 202.5 MHz 31P NMR. The 1H chemical shifts for the trans-syn thymine dimer unit of the decamer were found to be quite similar to those found for the trans-syn thymine dimer of TpT. Upon photolysis at 254 nm, CGTAT[t,s]TATGC was converted to a major product which coeluted with authentic CGTATTATGC and a minor product which coeluted with authentic CGTAT[c,s]TATGC, further supporting the presence of an intact trans-syn thymine dimer unit.
描述了一种通过固相DNA合成技术将反式-顺式胸腺嘧啶二聚体序列特异性引入寡核苷酸的构建模块的合成方法。通过部分自动化程序合成了CGTAT[t,s]TATGC，总产率为48%。添加反式-顺式胸腺嘧啶二聚体构建模块的逐步偶联产率为58%。含有二聚体的寡核苷酸通过500 MHz 1H COSY和NOESY光谱以及202.5 MHz 31P NMR进行表征。发现十聚体中反式-顺式胸腺嘧啶二聚体单元的1H化学位移与TpT的反式-顺式胸腺嘧啶二聚体的化学位移非常相似。在254 nm光解时，CGTAT[t,s]TATGC转化为与真实的CGTATTATGC共洗脱的主要产物和与真实的CGTAT[c,s]TATGC共洗脱的次要产物，进一步支持了完整的反式-顺式胸腺嘧啶二聚体单元的存在。
