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GST fusion expression and purification of human NDRG1
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 20
- DOI:
- 10.3321/j.issn:1000-2790.2005.20.004
- Key Word:
- NDRG1;GST;基因表达;谷胱甘肽琼脂糖-4B
Abstract： 目的: 为了更好的了解NDRG1的功能,通过pull-down技术寻找与NDRG1相互作用的蛋白. 方法: 从已构建好的NDRG1-pPROEXHTb重组载体中酶切下NDRG1 cDNA,插入融合表达载体pGEX-4T-1,序列测定后,转化DH-5α感受态细胞,在含氨苄青霉素的LB中高效表达. 表达的GST-NDRG1经过谷胱甘肽琼脂糖-4B亲和层析纯化,并与高表达细胞系HHCC孵育,通过SDS-PAGE检测和NDRG1相互作用的蛋白. 结果: 纯化蛋白和HHCC孵育后,在Mr 4.0×104处出现新增条带,应是与NDRG1相互作用的未知蛋白. 结论: 在肿瘤细胞HHCC中存在与NDRG1相互作用的蛋白.
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