Kikuchi Y, Sakaguchi K
Nucleic Acids Res. 1978 Feb;5(2):591-8. doi: 10.1093/nar/5.2.591.
The oligoribonucleotide, A-A-A-C-U-U-U-Gp, constituting a segment of RNA bacteriophage Qbeta coat protein gene was efficiently synthesized at a milligram scale by a combination of enzymatic methods using bacteriophage T4 RNA ligase and the thermophilic polynucleotide phosphorylase. A-A-A-Cp was synthesized from A-A-A and pCp by the newly developed mononucleotide addition method using T4 RNA ligase in a yield of 83%, followed by dephosphorylation with bacterial alkaline phosphatase to obtain A-A-A-C. pU-U-U-Gp was synthesized from pU-U-U and GDP by the simultaneous action of polynucleotide phosphorylase and RNase T1 in a yield of 32%. finally, the two oligonucleotides (A-A-A-C and pU-U-U-Gp) were ligated with T4 RNA ligase and the octanucleotide, A-A-A-C-U-U-U-Gp, was obtained in a yield of 85%.
构成RNA噬菌体Qβ外壳蛋白基因片段的寡核糖核苷酸A - A - A - C - U - U - U - Gp，通过使用噬菌体T4 RNA连接酶和嗜热多核苷酸磷酸化酶的酶法组合，以毫克规模高效合成。利用T4 RNA连接酶通过新开发的单核苷酸添加方法，由A - A - A和pCp合成A - A - A - Cp，产率为83%，随后用细菌碱性磷酸酶进行去磷酸化以获得A - A - A - C。通过多核苷酸磷酸化酶和RNase T1的同时作用，由pU - U - U和GDP合成pU - U - U - Gp，产率为32%。最后，将两个寡核苷酸（A - A - A - C和pU - U - U - Gp）用T4 RNA连接酶连接，得到八核苷酸A - A - A - C - U - U - U - Gp，产率为85%。
