Kikuchi Y, Hirai K, Sakaguchi K
J Biochem. 1975 Feb;77(2):469-72. doi: 10.1093/oxfordjournals.jbchem.a130747.
A method was developed for stepwise wynthesis of oligonucleotides of difined wequence using 2'(3')-O-dihydrocinnamoyl-nucleoside 5'-diphosphates as substrates for polynucleotide phosphorylase [ED 2.7.7.8]. Polynucleotide phosphorylase from Thermus thermophilus catalyzed the transfer of one 2'(3')-blocked ADP to the 3'-terminus of the primer trinucleoside diphosphate, ApApA. The product was 2'(3')-substituted triadenylyladenosine. The blocking group, dihydrocinnamoyl, could be removed completely from the product without destruction of the phosphodiester bond using alpha-chymotrypsin [ED 3.4.21.1] at neutral pH.
开发了一种使用2'(3')-O-二氢肉桂酰核苷5'-二磷酸作为多核苷酸磷酸化酶[EC 2.7.7.8]的底物逐步合成特定序列寡核苷酸的方法。嗜热栖热菌的多核苷酸磷酸化酶催化一个2'(3')-封闭的ADP转移到引物三磷酸腺苷二磷酸ApApA的3'-末端。产物是2'(3')-取代的三腺苷酰腺苷。在中性pH条件下,使用α-胰凝乳蛋白酶[EC 3.4.21.1]可以从产物中完全去除封闭基团二氢肉桂酰,而不会破坏磷酸二酯键。