Dumont J, Legrain M, Portetelle D, Brasseur R, Burny A, Hilger F
Unit of Microbiology, Faculty of Agronomy, Gembloux, Belgium.
Gene. 1989 Jul 15;79(2):219-26. doi: 10.1016/0378-1119(89)90204-7.
Bovine leukemia virus (BLV) p24 gene was expressed in Saccharomyces cerevisiae under the control of the PHO5 (encoding repressible acid phosphatase, rAPase) promoter. Yeast cells were transformed by a yeast-E. coli shuttle vector carrying the PHO5 promoter, the p24 gene and the CYC1 transcription terminator. After low inorganic phosphate (Pi) induction of the PHO5 promoter, p24 accumulated in the producing cells up to a concentration representing 10% of total soluble proteins. The expression level of p24 gene was not increased by insertion of the positive regulatory gene PHO4 on the p24 expression vector. The p24 produced in this system and incubated in crude yeast extract showed a remarkably high resistance to proteolytic degradation, a feature that presumably correlates with the compact globular conformation of the protein combined to the stabilizing effect of the N-terminal residue.
牛白血病病毒（BLV）的p24基因在酿酒酵母中于PHO5（编码可阻遏酸性磷酸酶，rAPase）启动子的控制下表达。酵母细胞通过携带PHO5启动子、p24基因和CYC1转录终止子的酵母-大肠杆菌穿梭载体进行转化。在低无机磷酸盐（Pi）诱导PHO5启动子后，p24在产生细胞中积累，浓度达到总可溶性蛋白的10%。通过在p24表达载体上插入正调控基因PHO4，p24基因的表达水平并未增加。在该系统中产生并在粗酵母提取物中孵育的p24对蛋白水解降解表现出显著的高抗性，这一特性可能与蛋白质紧密的球状构象以及N端残基的稳定作用相关。
