| 摘要: |
| β2微球蛋白（β2 m）是主要组织相容性复合体（MHC）-Ⅰ类分子的轻链部分，该蛋白的原核表达与纯化是制备MHC-Ⅰ类分子的首要条件。利用已经构建好的β2m原核表达载体pET23a＋β2 m，在大肠杆菌（E.coli）中获得稳定表达。原核表达的β2 m大部分在包涵体中，包涵体蛋白经充分洗涤、变性（尿素溶解）和复性，并以强阴离子交换柱层析（Q-Sepharose）纯化，获得SDS-PAGE纯的人重组β2 m，Western印迹法分析表明，该蛋白具有与抗人天然β2 m抗体反应的特性。 |
| 关键词: β2微球蛋白 重组蛋白 包涵体 复性 |
| DOI： |
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| 基金项目:国家自然科学基金项目（30370383） |
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| Human β2-microglobulin expression and purification in Escherichia coli |
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| Abstract: |
| Human β2-microglobulin (β2m) is a light chain of major histocompatibility complex (MHC) class Ⅰ molecule.Expression and purification of this protein in Escherichia coli (E.coli) is a prerequisite to the preparation of MHC-Ⅰ molecule.The expression vector pET23a+β2m was presented，in which the complete sequence of β2m gene was inserted.Constant-yield expression of β2m was achieved in E.coli transformed with the expression vector，and most of the recombinant β2m existed in the inclusion body after IPTG induction.The inclusion body was washed extensively and β2m in the inclusion body was solublized with 6 M urea.The β2m was refolded by dialysis and purified by ion-exchange chromatography (Q-Sepharose).Western blotting assay indicated that the polyclonal antibody against human native β2m could react specifically with the recombinant protein.The purified protein appeared as a single band on both SDS-PAGE and Western blotting，indicating that it was chemical and antigenic pure.This work provides the basis for the further study of MHC-Ⅰ molecule. |
| Key words: β2-microglobulin recombinant protein inclusion body refolding |
