Kramer R A, DeChiara T M, Schaber M D, Hilliker S
Proc Natl Acad Sci U S A. 1984 Jan;81(2):367-70. doi: 10.1073/pnas.81.2.367.
The promoter/regulator region from the yeast repressible acid phosphatase gene was used to construct a vector for the regulated expression of cloned genes in yeast. The gene for human leukocyte interferon was inserted into this vector. Yeast cells transformed with the resulting plasmid produced significant amounts of interferon only when grown in medium lacking inorganic phosphate. Mutants in two acid phosphatase regulatory genes (coding for a defective repressor and a temperature-sensitive positive regulator) were used to develop a yeast strain that grew well at a high temperature (35 degrees C) but produced interferon only at a low temperature (23 degrees C), independent of phosphate concentration.
利用酵母可阻遏酸性磷酸酶基因的启动子/调控区构建了一个载体,用于在酵母中调控克隆基因的表达。将人白细胞干扰素基因插入该载体。用所得质粒转化的酵母细胞只有在缺乏无机磷酸盐的培养基中生长时才会产生大量干扰素。利用两个酸性磷酸酶调控基因的突变体(编码缺陷型阻遏物和温度敏感型正调控因子)培育出一种酵母菌株,该菌株在高温(35℃)下生长良好,但仅在低温(23℃)下产生干扰素,且与磷酸盐浓度无关。