重组人IL-10在毕赤酵母X-33及SMD1168中表达效率的比较

目的研究重组人IL-10（rhIL-10）在两株毕赤酵母X-33和SMD1168的表达水平,为rhIL-10的表达选择合适的表达菌株。方法以0.5%甲醇分别诱导毕赤酵母X-33和SMD1168表达rhIL-10,用SDS-PAGE和WesternBlot分析和鉴定rhIL-10,用ELISA法测定培养液rhIL-10含量,用Bradford法测定培养液总蛋白含量并计算rhIL-10占总蛋白的比例。比较同等培养条件下两株酵母表达rhIL-10和总蛋白的水平。结果 SDS-PAGE和Western Blot均检测到42kD左右的蛋白条带;rhIL-10在X-33中的表达量平均达20.00mg/L,在SMD1168中的表达量平均达1.90mg/L,在两株不同酵母中的表达量有显著性差异（P〈0.05）;同时段的X-33培养液中总蛋白质含量平均为50.00mg/L,rhIL-10占总蛋白的40.00%;SMD1168培养液中总蛋白质含量平均为48.23mg/L,rhIL-10占总蛋白的4.00%。结论 rhIL-10在不同的毕赤酵母表达效率有差异,X-33较1168更适于rhIL-10的表达。

Expression of recombinant human interleukin 10 in X-33 and SMD1168 Pi-chia pastoris and comparison of the efficiency

Objective The current study was aim to explore the level of of recombinant human interleukin 10(rhIL-10) expressing in X-33 and SMD1168 Pichia pastoris and choose a suitable host for expression of rhIL10.Methods rhIL-10 was expressed in Pichia pastoris X-33 and SMD1168 Pichia pastoris by inducing with methanol 0.5%.The analysis and identification were carried out by SDS-PAGE and Western Blot.The content of rhIL-10 in culture solution was determined by ELISA,and the content of total protein in culture soluti...