Kravchenko V V, Gileva I P, Shamin V V, Kulichkov V A, Dobrynin V N
Bioorg Khim. 1987 Sep;13(9):1176-85.
Using gene fragments encoding the leader peptide of E. coli tryptophane operon (as duplicated fragment HhaI-140) or M13 phage coat protein (as TaqI-381 or HaeIII-1623 fragments) and basing on pDS1 family of plasmids, expression vectors have been constructed which contained transcription promoters Ptrp, PVIII, and Pv + PVIII, respectively. An artificial gene for human leukocyte interferon alpha 2 (ifn-alpha 2) has been cloned into these plasmids, so that its transcription was a part of polycistronic mRNA and preceding translation was terminated upstream to the ribosome binding site and starting codon of the interferon gene. E. coli cells harbouring these recombinant plasmids provided high level of the interferon biosynthesis. The effect of the mRNA length on the amount of protein synthesised under control of the M13 coat protein transcription-translation signals has been found.
利用编码大肠杆菌色氨酸操纵子前导肽的基因片段(如重复片段HhaI - 140)或M13噬菌体外壳蛋白的基因片段(如TaqI - 381或HaeIII - 1623片段),并基于pDS1质粒家族构建了表达载体,这些载体分别包含转录启动子Ptrp、PVIII和Pv + PVIII。将人白细胞干扰素α2(ifn - α2)的人工基因克隆到这些质粒中,使其转录成为多顺反子mRNA的一部分,且在干扰素基因的核糖体结合位点和起始密码子上游终止翻译。携带这些重组质粒的大肠杆菌细胞实现了高水平的干扰素生物合成。还发现了mRNA长度对在M13外壳蛋白转录 - 翻译信号控制下合成的蛋白量的影响。