Construction of a high sensitive escherichia coli alkaline phosphatase reporter system for screening affinity peptides
文献类型：期刊论文
| 作者 | Huang, Xu; Zhang, Xian-En; Zhou, Ya-Feng; Zhang, Zhi-Ping; Cass, Anthony E. G. |
| 刊名 | Journal of biochemical and biophysical methods
 |
| 出版日期 | 2007-04-10 |
| 卷号 | 70期号:3页码:435-439 |
| 关键词 | E. coli alkaline phosphatase Fusion protein Affinity peptide Screening system |
| ISSN号 | 0165-022X |
| DOI | 10.1016/j.jbbm.2006.10.006 |
| 通讯作者 | Zhang, xian-en(zhangxe@most.cn) |
| 英文摘要 | An enzyme-linker-peptide fusion protein reporter system was constructed for sensitive analysis of affinity of peptide ligands to their receptor. an e. coli alkaline phosphatase (eap) mutant enzyme with high catalytic activity was selected as the reporter protein. interaction of affinity peptide and streptavidin was applied as demonstration of the method. three affinity peptides, strep-tag i (si), strep-tag ii (sii) and streptavidin binding peptide (sbp) were genetically fused to the c-terminal of eap respectively, with an insertion of a flexible linker peptide in between. the enzyme activity of the eap fusions showed no obvious change. after expression and purification, the eap-affinity peptide fusions were applied to the streptavidin modified surface. binding of the fusions to the surface through interaction of affinity peptides to streptavidin was indicated by color generated from conversion of the substrate by ear the relative affinity and specificity of each affinity peptides to the immobilized streptavidin were then evaluated with high sensitivity and broad detection range. this method may be used for effective high-throughput screening of high affinity peptide from the peptide pool. (c) 2006 elsevier b.v. all rights reserved. |
| WOS关键词 | SURFACE DISPLAY ; PROTEIN-PURIFICATION ; RECOMBINANT PROTEINS ; BINDING PEPTIDES ; RIBOSOME DISPLAY ; FUSION PROTEINS ; STREPTAVIDIN ; LIBRARIES ; PHAGE ; TAG |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biophysics |
| WOS类目 | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biophysics |
| 语种 | 英语 |
| WOS记录号 | WOS:000245426400015 |
| 出版者 | ELSEVIER SCIENCE BV |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2375345 |
| 专题 | 武汉病毒研究所 |
| 通讯作者 | Zhang, Xian-En |
| 作者单位 | 1.Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China 2.Univ London Imperial Coll Sci Technol & Med, Inst Biomed Engn, London SW7 2AY, England 3.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China |
| 推荐引用方式 GB/T 7714 | Huang, Xu,Zhang, Xian-En,Zhou, Ya-Feng,et al. Construction of a high sensitive escherichia coli alkaline phosphatase reporter system for screening affinity peptides[J]. Journal of biochemical and biophysical methods,2007,70(3):435-439. |
| APA | Huang, Xu,Zhang, Xian-En,Zhou, Ya-Feng,Zhang, Zhi-Ping,&Cass, Anthony E. G..(2007).Construction of a high sensitive escherichia coli alkaline phosphatase reporter system for screening affinity peptides.Journal of biochemical and biophysical methods,70(3),435-439. |
| MLA | Huang, Xu,et al."Construction of a high sensitive escherichia coli alkaline phosphatase reporter system for screening affinity peptides".Journal of biochemical and biophysical methods 70.3(2007):435-439. |
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来源：武汉病毒研究所
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