A MutS-based protein chip for detection of DNA mutations
文献类型：期刊论文
| 作者 | Bi, LJ; Zhou, YF; Zhang, XE; Deng, JY; Zhang, ZP; Xie, B; Zhang, CG; Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China |
| 刊名 | ANALYTICAL CHEMISTRY
 |
| 出版日期 | 2003 |
| 卷号 | 75期号:16页码:4113-4119 |
| ISSN号 | 0003-2700 |
| 英文摘要 | This paper describes a new protein chip method for detection of single-base mismatches and unpaired bases of DNA, using a genetic fusion molecular system Trx-His(6)-Linker peptide-Strep-tagII-linker peptide-MutS (THLSLM). The THLSLM coding sequence was constructed by attaching Strep-tag 11 and mutS gene to pET32a (+) sequentially with insertion of a linker peptide coding sequence before and behind Strep-tagII gene, respectively. THLSLM was expressed in E. coli AD494 (DE3) and purified using Ni2+-chelation affinity resin. THLSLM retained both mismatch recognition activity and streptavidin binding affinity. THLSLM was then immobilized on the chip matrix coated with streptavidin through the Strep-tag II-streptavidin binding reaction. The resulting protein chip was used to detect the mismatched and unpaired mutations in the synthesized oligonucleotides, as well as a single-base mutation in rpoB gene from Mycobacterium tuberculosis, with high specificity. The method could potentially serve as a platform to develop the high-throughput technology for screening and analysis of genetic mutations. |
| 学科主题 | Chemistry, Analytical |
| 语种 | 英语 |
| WOS记录号 | WOS:000184798100020 |
| 公开日期 | 2011-09-23 |
| 源URL | [http://210.72.129.5/handle/321005/55379]  |
| 专题 | 沈阳应用生态研究所_沈阳应用生态研究所 |
| 通讯作者 | Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China |
| 推荐引用方式 GB/T 7714 | Bi, LJ,Zhou, YF,Zhang, XE,et al. A MutS-based protein chip for detection of DNA mutations[J]. ANALYTICAL CHEMISTRY,2003,75(16):4113-4119. |
| APA | Bi, LJ.,Zhou, YF.,Zhang, XE.,Deng, JY.,Zhang, ZP.,...&Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China.(2003).A MutS-based protein chip for detection of DNA mutations.ANALYTICAL CHEMISTRY,75(16),4113-4119. |
| MLA | Bi, LJ,et al."A MutS-based protein chip for detection of DNA mutations".ANALYTICAL CHEMISTRY 75.16(2003):4113-4119. |
入库方式： OAI收割
来源：沈阳应用生态研究所
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