Engineered SNAP-MBD2b proteins for specific recognition of methylated DNA
文献类型：期刊论文
| 作者 | Zhang DaPeng
|
| 刊名 | SCIENCE CHINA-CHEMISTRY
 |
| 出版日期 | 2014 |
| 卷号 | 57期号:7页码:1019-1025 |
| 关键词 | methylated DNA MBD2b SNAP-MBD2b SNAP-tag histidine-tagged affinity chromatography (AC) cation exchange chromatography (CXC) |
| ISSN号 | 1674-7291 |
| 中文摘要 | Methyl-CpG-binding domain (MBD) proteins can specifically recognize and bind methylated CpG sites of DNA, thus repress gene transcription. In this study, we designed and expressed two recombinant proteins, MBD2b and SNAP-MBD2b, in E. coli. An optimized protocol was developed to purify the proteins using Ni-NTA affinity cartridge and cation exchange resin. The engineered proteins purified by this method exhibited more than 93% purity and high binding avidity. We found that both SNAP-MBD2b and MBD2b were prone to aggregate during dialysis. However, this could be prevented by the use of 0.3 mol/L NaCl. The fusion of SNAP-tag with MBD2b significantly enhanced the expression of MBD2b protein in E. coli and reduced the adsorption of MBD2b on solid interfaces involved in protein purification and immobilization. The engineered proteins can be used for the study of interaction with methylated DNA and the assays for DNA methylation. |
| WOS记录号 | WOS:000338450800014 |
| 公开日期 | 2015-03-24 |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/9359]  |
| 专题 | 生态环境研究中心_环境化学与生态毒理学国家重点实验室 |
| 推荐引用方式 GB/T 7714 | Zhang DaPeng. Engineered SNAP-MBD2b proteins for specific recognition of methylated DNA[J]. SCIENCE CHINA-CHEMISTRY,2014,57(7):1019-1025. |
| APA | Zhang DaPeng.(2014).Engineered SNAP-MBD2b proteins for specific recognition of methylated DNA.SCIENCE CHINA-CHEMISTRY,57(7),1019-1025. |
| MLA | Zhang DaPeng."Engineered SNAP-MBD2b proteins for specific recognition of methylated DNA".SCIENCE CHINA-CHEMISTRY 57.7(2014):1019-1025. |
入库方式： OAI收割
来源：生态环境研究中心
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