Construction and characterization of different muts fusion proteins as recognition elements of dna chip for detection of dna mutations
文献类型：期刊论文
| 作者 | Bi, LJ; Zhou, YF; Zhang, XE; Deng, JY; Wen, JK; Zhang, ZP |
| 刊名 | Biosensors & bioelectronics
 |
| 出版日期 | 2005-07-15 |
| 卷号 | 21期号:1页码:135-144 |
| 关键词 | Muts fusion proteins Dna chip Mutation detection |
| ISSN号 | 0956-5663 |
| DOI | 10.1016/j.bios.2004.08.045 |
| 通讯作者 | Zhang, xe() |
| 英文摘要 | Three muts fusion systems were designed as the mutation recognition and signal elements of dna chips for detection of dna mutations. the expression vectors containing the encoding sequences of three recombinant proteins, trx-his(6)-gfp-(ser-gly)(6)-muts (thglm), trx-his(6)-(ser-gly)(6)-strep tagii-(ser-gly)(6)-muts (thlslm) and trx-his(6)-(ser-gly)(6)-muts (thlm), were constructed by gene slicing in vitro. thglm, thlslm and thlm were then expressed in escherichia coli ad494(de3), respectively. sds-page analysis revealed that each of the expected proteins was similar to 30% of the total bacterial proteins. the recombinant proteins were purified to the purity over 90% by immobilized metal (co2+) chelation affinity chromatography. bioactivity assay indicated that three fusion proteins retained the mismatch-binding activity and the functions of other fusion partners. dna chips arrayed both mismatched and unpaired dna oligonucleotides as well as rpob gene from mycobacterium tuberculosis were prepared. thglm, thlslm and thlm that was labeled with fluorolink (tm) cy3 reactive dye, were then used as both mutation recognition and labeling elements of dna chips. the resulting dna chips were used to detect the mismatched and unpaired mutations in the synthesized oligonucleotides and single base mutation in rpob gene of m. tuberculosis that is resistant to rifamycin. (c) 2004 elsevier b.v. all rights reserved. |
| WOS关键词 | MISMATCH BINDING-PROTEIN ; GRADIENT GEL-ELECTROPHORESIS ; POINT MUTATIONS ; MYCOBACTERIUM-TUBERCULOSIS ; HETERODUPLEX DNA ; REPAIR ; GENE ; SYSTEM |
| WOS研究方向 | Biophysics ; Biotechnology & Applied Microbiology ; Chemistry ; Electrochemistry ; Science & Technology - Other Topics |
| WOS类目 | Biophysics ; Biotechnology & Applied Microbiology ; Chemistry, Analytical ; Electrochemistry ; Nanoscience & Nanotechnology |
| 语种 | 英语 |
| WOS记录号 | WOS:000232268900016 |
| 出版者 | ELSEVIER ADVANCED TECHNOLOGY |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2375254 |
| 专题 | 武汉病毒研究所 |
| 通讯作者 | Zhang, XE |
| 作者单位 | 1.Chinese Acad Sci, Inst Biophys, Beijing 100101, Peoples R China 2.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. Construction and characterization of different muts fusion proteins as recognition elements of dna chip for detection of dna mutations[J]. Biosensors & bioelectronics,2005,21(1):135-144. |
| APA | Bi, LJ,Zhou, YF,Zhang, XE,Deng, JY,Wen, JK,&Zhang, ZP.(2005).Construction and characterization of different muts fusion proteins as recognition elements of dna chip for detection of dna mutations.Biosensors & bioelectronics,21(1),135-144. |
| MLA | Bi, LJ,et al."Construction and characterization of different muts fusion proteins as recognition elements of dna chip for detection of dna mutations".Biosensors & bioelectronics 21.1(2005):135-144. |
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来源：武汉病毒研究所
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