Airenne K J, Sarkkinen P, Punnonen E L, Kulomaa M S
Department of Biology, University of Jyväskylä, Finland.
Gene. 1994 Jun 24;144(1):75-80. doi: 10.1016/0378-1119(94)90206-2.
A recombinant avidin (re-Avd), containing amino acids (aa) 1-123 of the native chicken egg-white Avd, was produced in Escherichia coli. When cells were grown at 37 degrees C production was over 1 microgram/ml, due to altering the codon preference of the first ten codons. The re-Avd was recovered as a soluble protein from cells grown at 25 or 30 degrees C, whereas at 37 degrees C it was mostly insoluble in inclusion bodies. Our results indicated that, despite the potentially harmful biotin-binding activity of Avd, it is possible to produce biologically active Avd in E. coli which then can easily be purified by affinity chromatography on a biotin column in a single step.
一种重组抗生物素蛋白（re-Avd），包含天然鸡蛋白抗生物素蛋白的第1至123个氨基酸，在大肠杆菌中产生。当细胞在37℃生长时，由于改变了前十个密码子的密码子偏好性，产量超过1微克/毫升。重组抗生物素蛋白从在25或30℃生长的细胞中以可溶性蛋白形式回收，而在37℃时它大多不溶于包涵体。我们的结果表明，尽管抗生物素蛋白具有潜在有害的生物素结合活性，但仍有可能在大肠杆菌中产生具有生物活性的抗生物素蛋白，然后通过在生物素柱上进行一步亲和色谱法即可轻松纯化。
