Brinkmann U, Mattes R E, Buckel P
Department of Genetics, Boehringer Mannheim GmbH, Penzberg (F.R.G.).
Gene. 1989 Dec 21;85(1):109-14. doi: 10.1016/0378-1119(89)90470-8.
We have observed that proteins, such as human tissue-type plasminogen activator, pro-urokinase or gp41 of human immunodeficiency virus, which have a high content of rare codons in their respective genes, are not readily expressed in Escherichia coli. Furthermore induction of these heterologous genes leads to growth inhibition and plasmid instability. Supplementation with tRNA(AGA/AGG(Arg)) by cotransfection with the dnaY gene, which supplies this minor tRNA, resulted in high-level production with greatly improved cell viability and plasmid stability.
我们观察到，诸如人组织型纤溶酶原激活剂、尿激酶原或人类免疫缺陷病毒的gp41等蛋白质，其各自基因中稀有密码子含量很高，在大肠杆菌中不易表达。此外，诱导这些异源基因会导致生长抑制和质粒不稳定。通过与提供这种次要tRNA的dnaY基因共转染来补充tRNA(AGA/AGG(Arg))，可实现高水平表达，并大大提高细胞活力和质粒稳定性。
