重组人Tau蛋白的表达、纯化及鉴定

Expression, Purification and Identification of Recombinant Human Tau Pro-tein

目的 重组人Tau蛋白的诱导表达和纯化。方法 将携带 Tau蛋白基因的工程菌株经IPTG诱导表达后,通过超声破碎、硫酸按盐析、离子交换层析、电洗脱等方法纯化Tau蛋白,以SDS-PAGE、非变性PAGE、Westem blot和N末端氨基酸测定等方法进行鉴定。结果 纯化Tau蛋白的得率为28.5％,在SDS-PAGE和非变性PAGE上呈均一的蛋白条带,其相对分子质量约为59000,N末端5个氨基酸测定结果为NH2-Met-Ala-Glu-Pro-Arg-。结论 本实验采用纯化重组人Tau蛋白方法是可行的。

Objective To express and purify recombinant human Tau protein. Methods The recom-binant bacterial strain containing Tau gene was induced with IPTG, and the expressed product was purified by sonication, salting out with ammonium sulfate, ion exchange charomatography and electric elution, then purified by SDS - PAGE, non - denaturing PAGE, Western blot and amino acid sequencing of N - terminal. Results The recovery rate of purified Tau protein was 28.5% . Either SDS - PAGE or non - denaturing PAGE profile showed a single protein band with a relative molecular weight of 59000. The sequence of 5 amino acids at the N - terminal of the protein was NH2 - Met - Ala - Glu - Pro - Arg - . Conclusion The methods in this paper were effective in the purification of recombinant human Tau protein. It laid a foundation of study on pathogene-sis and early diagnosis of Alzheimer's disease.