Journal of Capital Medical University ›› 2011, Vol. 32 ›› Issue (6): 798-801.doi: 10.3969/j.issn.1006-7795.2011.06.018

• 帕金森病的发病机制研究 • Previous Articles     Next Articles

Prokaryotic expression and purification of recombinant human SORL1 protein

LI Shuang1, LI Yao-hua1, YE Yi-wen1, LI Xin1, YU Shun1, YANG Hui2, CHEN Biao1   

  1. 1. Department of Neurobiology, Beijing Institute of Geriatrics, Key Laboratory for Neurodegenerative Diseases, Ministry of Education;Xuanwu Hospital, Capital Medical University, Beijing 100053, China;2. Beijing institute for Neuroscience, Capital Medical University, Beijing center for Neural Regeneration & Repair;Key Laboratory for Neurodegenerative Disease, Ministry of Education, Beijing 100069, China
  • Received:2011-10-16 Revised:1900-01-01 Online:2011-12-21 Published:2011-12-21

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Abstract: Objective To prepare the recombinant human SORL1 protein, and provide the experimental foundation for study about the pathogenesis of Alzheimer's disease. Methods A human sorl1 cDNA fragment(5 923~6 260 bp) was amplified by PCR, and subucloned into pET-28a(+) vector. Recombinant plasmid was expressed by E.coli BL21(DE3), and the recombinant protein was purified by liquid chromatography. Results A 358 bp cDNA fragment was amplified by PCR method. Its structure between the ATG and TGA was completely consistent with the human sorl1 cDNA fragment(5 923~6 260 bp). The recombinant plasmid in E.coli BL21(DE3) was highly expressed and its expression product was mainly in the inclusion body. The purified protein on SDS-PAGE demonstrated a single band, which appeared to have a molecular weight of about 13 000. Conclusion Human sorl1 cDNA fragment was highly expressed in prokaryotic cells. The purified recombinant protein can be used to prepare antibodies.

Key words: sortilin-related receptor, L(DLR class) A repeats-containing, recombinant protein, Alzheimer's disease

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