Kravchenko V V, Gileva I P, Shamin V V, Kulichkov V A, Dobrynin V N
Bioorg Khim. 1987 Sep;13(9):1186-93.
Using a chemically synthesised adapter, the coding part of an artificial gene for human leukocyte alpha 2 interferon (ifn-alpha 2) has been duplicated. The adapter contained a termination signal of the first gene (TAA) within the Shine-Dalgarno sequence of the second gene (TAAGGA), distance between the terminating codon and starting codon of the second gene being 11 nucleotides. In another case this distance was 69 nucleotides, with the same SD sequence. The expression of the tandems as a part of polycistrons has been studied under control of promoters Plac, (Ptrp)2 of E. coli, and PVIII of M13 phage. It was found that tandems of ifn-alpha 2 genes in polycistronic structures trp L-ifn-ifn and IX-VIII-ifn-ifn under control of promoters (Ptrp)2 and PVIII, respectively, provided high level of the interferon biosynthesis, thus differing from the tandem under Plac promoter control, which had only ifn-ifn translation coupling.
利用化学合成的衔接子,人白细胞α2干扰素(ifn-α2)人工基因的编码部分已被复制。该衔接子在第二个基因的Shine-Dalgarno序列(TAAGGA)内包含第一个基因的终止信号(TAA),第二个基因的终止密码子与起始密码子之间的距离为11个核苷酸。在另一种情况下,该距离为69个核苷酸,SD序列相同。作为多顺反子一部分的串联体的表达已在大肠杆菌的启动子Plac、(Ptrp)2以及M13噬菌体的PVIII控制下进行了研究。结果发现,分别在启动子(Ptrp)2和PVIII控制下的多顺反子结构trp L-ifn-ifn和IX-VIII-ifn-ifn中的ifn-α2基因串联体,可提供高水平的干扰素生物合成,这与Plac启动子控制下的串联体不同,后者只有ifn-ifn翻译偶联。