编码聚（L-天冬氨酰-L-苯丙氨酸）的合成重复多聚基因在大肠杆菌中的表达

Doel M T, Eaton M, Cook E A, Lewis H, Patel T, Carey N H

Nucleic Acids Res. 1980 Oct 24;8(20):4575-92. doi: 10.1093/nar/8.20.4575.

Two dodecadeoxynucleotides of defined sequence have been synthesised by phosphotriester methodology. They can be polymerised to give a double stranded DNA which codes, when read in the correct phase, for the repeating dipeptide poly(aspartyl-phenylalanine). This polymeric DNA has been cloned in E. coli K12 using as vector a plasmid having a controllable bacterial promoter upstream of the insertion site. Clones containing genes coding for up to 150 repeats of (aspartyl-phenylalanine) have been isolated and characterised. The polymeric inserts appear to be stable over many generations and are expressed in E. coli under the control of the bacterial promoter, to give a polymer of phenylalanine and aspartic acid which may be broken down enzymically to yield aspartyl-phenylalanine.

通过磷酸三酯法合成了两个具有特定序列的十二聚脱氧核苷酸。它们可以聚合形成双链DNA，当以正确的相位读取时，该双链DNA编码重复的二肽聚（天冬氨酰 - 苯丙氨酸）。使用在插入位点上游具有可控细菌启动子的质粒作为载体，将这种聚合DNA克隆到大肠杆菌K12中。已经分离并表征了含有编码多达150个（天冬氨酰 - 苯丙氨酸）重复序列的基因的克隆。聚合插入片段在许多代中似乎是稳定的，并在细菌启动子的控制下在大肠杆菌中表达，产生苯丙氨酸和天冬氨酸的聚合物，该聚合物可被酶分解产生天冬氨酰 - 苯丙氨酸。