Sleep D, Belfield G P, Ballance D J, Steven J, Jones S, Evans L R, Moir P D, Goodey A R
Delta Biotechnology Limited, Nottingham, Great Britain.
Biotechnology (N Y). 1991 Feb;9(2):183-7. doi: 10.1038/nbt0291-183.
We describe a system that facilitates the selection of host mutants that overproduce a range of secreted and internally produced heterologous proteins in Saccharomyces cerevisiae. These mutants were initially selected for their ability to oversecrete recombinant human albumin (rHA), as detected by a direct visual assay that relies upon antibody precipitation in solid media. Yeast strains that were able to synthesize and secrete increased levels of rHA also produced elevated levels of internally expressed proteins including alpha 1-antitrypsin Pittsburgh variant and plasminogen activator inhibitor type 2.
我们描述了一种系统，该系统有助于在酿酒酵母中选择能够过量产生一系列分泌型和胞内产生的异源蛋白的宿主突变体。这些突变体最初是因其过量分泌重组人白蛋白（rHA）的能力而被筛选出来的，这是通过一种直接视觉检测法检测到的，该检测法依赖于固体培养基中的抗体沉淀。能够合成并分泌更高水平rHA的酵母菌株也产生了更高水平的胞内表达蛋白，包括α1-抗胰蛋白酶匹兹堡变体和纤溶酶原激活物抑制剂2型。
