Khorana H G, Agarwal K L, Besmer P, Büchi H, Caruthers M H, Cashion P J, Fridkin M, Jay E, Kleppe K, Kleppe R, Kumar A, Loewen P C, Miller R C, Minamoto K, Panet A, RajBhandary U L, Ramamoorthy B, Sekiya T, Takeya T, van de Sande J H
J Biol Chem. 1976 Feb 10;251(3):565-70.
With the ultimate objective of the total synthesis of a tRNA gene including its transcriptional signals, an Escherichia coli tyrosine suppressor tRNA gene was chosen. The arguments in favor of this choice are presented. A plan for the total synthesis of the 126-nucleotide-long DNA duplex corresponding to a precursor (Altman S., and Smith, J. D. (1971) Nature New Biol. 233, 35) to the above tRNA is formulated. The plan involves: (a) the chemical synthesis of 26 deoxyribooligonucleotide segments, (b) polynucleotide ligase-catalyzed joining of several segments at a time to form a total of four DNA duplexes with appropriate comlementary single-stranded ends, and (c) the joining of the duplexes to form the entire DNA duplex. Ten accompanying papers describe the experimental realization of this objective.
为了实现包括转录信号在内的tRNA基因的全合成这一最终目标,选择了大肠杆菌酪氨酸抑制tRNA基因。文中阐述了支持这一选择的理由。制定了一个全合成126个核苷酸长的DNA双链体的计划,该双链体对应于上述tRNA的前体(阿尔特曼S.和史密斯J.D.(1971年)《自然新生物学》233, 35)。该计划包括:(a)化学合成26个脱氧核糖寡核苷酸片段,(b)多核苷酸连接酶催化一次连接几个片段,以形成总共四个具有适当互补单链末端的DNA双链体,以及(c)将双链体连接起来形成完整的DNA双链体。十篇附带论文描述了这一目标的实验实现情况。