Dittrich W, Williams K L, Slade M B
School of Biological Sciences, Macquarie University, Sydney, NSW, Australia.
Biotechnology (N Y). 1994 Jun;12(6):614-8. doi: 10.1038/nbt0694-614.
We have expressed useful amounts of three recombinant proteins in a new eukaryotic host/vector system. The cellular slime mold Dictyostelium discoideum efficiently secreted two recombinant products, a soluble form of the normally cell surface associated D. discoideum glycoprotein (PsA) and the heterologous protein glutathione-S-transferase (GST) from Schistosoma japonicum, while the enzyme beta-glucuronidase (GUS) from Escherichia coli was cell associated. Up to 20mg/l of recombinant PsA and 1mg/l of GST were obtained after purification from a standard, peptone based growth medium. The secretion signal peptide was correctly cleaved from the recombinant GST- and PsA-proteins and the expression of recombinant PsA was shown to be stable for at least one hundred generations in the absence of selection.
我们已在一种新型真核宿主/载体系统中表达了有用量的三种重组蛋白。细胞黏菌盘基网柄菌有效地分泌了两种重组产物，一种是通常与细胞表面相关的盘基网柄菌糖蛋白（PsA）的可溶性形式，另一种是来自日本血吸虫的异源蛋白谷胱甘肽 - S - 转移酶（GST），而来自大肠杆菌的β - 葡萄糖醛酸酶（GUS）则与细胞相关。从基于蛋白胨的标准生长培养基中纯化后，可获得高达20mg/l的重组PsA和1mg/l的GST。分泌信号肽已从重组GST和PsA蛋白上正确切割下来，并且在没有选择压力的情况下，重组PsA的表达显示至少在一百代内是稳定的。
