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目的探讨基于酵母的抗朊病毒药物筛选模型在初级筛选时的细胞培养最佳条件。方法通过表型分析的方法探讨了培养基pH值、初始菌液浓度、细胞生长时期3个因素对基于酵母的抗朊病毒药物筛选模型灵敏度的影响。结果培养基pH值能够影响酵母朊病毒[psi-]细胞的颜色表型,pH>7.46时,[psi-]酵母菌落变为白色,但对[PSI+]的聚集状态不造成影响;菌液初始浓度为A600nm=0.02时,相对于A600nm=0.5、A600nm=0.2时的药物治愈率要高几倍到几十倍;处于对数生长期的[PSI+]酵母细胞对药物的敏感性较差,而处于延滞期和稳定期的[PSI+]酵母细胞对药物的敏感性较强。结论酵母抗朊病毒药物筛选模型在初级筛选时的细胞培养最佳条件为pH<6,菌液初始浓度为A600nm=0.02,处于稳定期的酵母细胞。
OBJECTIVE: To study the best conditions for cell culture of yeast-based anti-prion drug screening model in primary screening. Methods The effects of pH, initial bacterial concentration and cell growth on the sensitivity of yeast-based anti-prion drug screening models were investigated by phenotypic analysis. Results The medium pH value could affect the color phenotype of yeast [psi-] cells. When pH> 7.46, the [psi-] yeast colony turned white but had no effect on the aggregation state of [PSI +]; initial bacterial broth The concentration of A600nm = 0.02, relative to the A600nm = 0.5, A600nm = 0.2 when the drug cure rate several times to several times higher; in the logarithmic growth phase [PSI +] Yeast cells less susceptible to drugs, and Yeast cells in [PSI +] lag phase and stationary phase are more drug sensitive. Conclusions The optimum condition of yeast cell culture in primary screening is pH <6, initial concentration of bacteria is A600nm = 0.02, and it is in stable yeast cells.