Chen D C, Chuang L T, Chen W P, Kuo T T
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, Republic of China.
Curr Genet. 1995 Feb;27(3):201-6. doi: 10.1007/BF00326149.
The toxicity of HBsAg in the secretion pathway of pep4 strains can be progressively reduced in modified SD media containing lower concentrations of ammonium sulphate. A procedure, combining a reduction of ammonium sulphate concentration in SD media with the disruption of the PEP4 gene of the host strain, was developed to enrich transformants which are not inhibited by HBsAg expressed in the secretion pathway. Abnormal growth of these non-inhibited transformants is characterized by the enlargement of cell morphology, a transition to pseudohyphal-like growth in nitrogen-starved media, an increase in HBsAg particle production, and the enhancement of growth rate in liquid media. This suggests a new approach to overcoming the toxicity of heterologous protein in the yeast secretion pathway.
在含有较低浓度硫酸铵的改良SD培养基中，pep4菌株分泌途径中HBsAg的毒性可逐渐降低。开发了一种将SD培养基中硫酸铵浓度降低与宿主菌株PEP4基因破坏相结合的方法，以富集不受分泌途径中表达的HBsAg抑制的转化体。这些未受抑制的转化体的异常生长表现为细胞形态增大、在氮饥饿培养基中向假菌丝样生长转变、HBsAg颗粒产量增加以及液体培养基中生长速率提高。这提示了一种克服酵母分泌途径中异源蛋白毒性的新方法。
