天然人α-1干扰素基因在大肠杆菌中的组成型表达。

Constitutive expression of a native human interferon-alpha 1 gene in Escherichia coli.

Ivanov I, Markova N, Bachvarov D, Alexciev K, Saraffova A, Maximova V, Tsaneva I, Markov G

Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia.

Int J Biochem. 1989;21(9):983-5. doi: 10.1016/0020-711x(89)90230-9.

Abstract

A plasmid for constitutive expression of the human interferon-alpha 1 (hIFN-alpha 1) gene in Escherichia coli is constructed on the basis of the cloning plasmid pBR322 using a strong synthetic promoter, synthetic ribosome binding site and a native hIFN-alpha 1 gene excised from a chromosomal clone. 2. The yield of recombinant hIFN-alpha 1 from E. coli LE392 cells transformed with the expression plasmid pJP1R9-hIFN-alpha 1 is evaluated to be 2-6 x 10(7) U/l bacterial culture for metabolic shaker and 6-8 x 10(7) U/l for fermentor.

摘要

基于克隆质粒pBR322构建了一种用于在大肠杆菌中组成型表达人α-1干扰素（hIFN-α1）基因的质粒，该质粒使用了一个强合成启动子、合成核糖体结合位点以及从染色体克隆中切除的天然hIFN-α1基因。2. 用表达质粒pJP1R9-hIFN-α1转化的大肠杆菌LE392细胞产生的重组hIFN-α1产量，对于代谢摇床培养的细菌培养物评估为2 - 6×10⁷ U/升，对于发酵罐培养为6 - 8×10⁷ U/升。