论文部分内容阅读
目的利用具有天然N-末端的重组鸡半胱氨酸蛋白酶抑制剂(chicken cystatin,cC)在毕赤酵母中高效分泌表达,研究淀粉样沉淀倾向。方法通过在巴氏毕赤酵母(Pichia pastoris)菌株X-33中甲醇诱导表达cC能与刚果红特异性结合特性和硫磺素T荧光分析聚积方法研究聚集的特性。结果重组cC形成的类淀粉样蛋白的聚集量有明显增加,30 d时,紫外分光光度检测,重组cC为1.21μmol/L,重组cC荧光强度由1.08增至18.05,即聚集程度增加。结论野生型cystatin C也有形成淀粉样沉淀倾向。
OBJECTIVE: To study the amyloid deposition tendency of recombinant chicken chicken cystatin (cC) with high natural N-terminal secretion in Pichia pastoris. Methods Aggregation was studied by methanol-induced expression of cC in Pichia pastoris strain X-33 with specific binding to Congo red and thioflavin T fluorescence analysis. Results The aggregation of amyloid protein formed by recombinant cC increased obviously. At 30 days, the fluorescence intensity of recombinant cC increased from 1.08 to 18.05 by UV spectrophotometry, the recombinant cC was 1.21 μmol / L, and the aggregation degree increased. Conclusion Wild type cystatin C also has the tendency to form amyloid deposits.