A simplified method for reconstituting active e. coli dna polymerase iii
文献类型：期刊论文
| 作者 | Lin, Shi-Qiang1,3; Bi, Li-Jun1; Zhang, Xian-En2 |
| 刊名 | Protein & cell
 |
| 出版日期 | 2011-04-01 |
| 卷号 | 2期号:4页码:303-307 |
| 关键词 | E. coli Dna polymerase iii Coexpression Purification |
| ISSN号 | 1674-800X |
| DOI | 10.1007/s13238-011-1032-3 |
| 通讯作者 | Bi, li-jun(blj@sun5.ibp.ac.cn) |
| 英文摘要 | Genome duplication in e. coli is carried out by dna polymerase iii, an enzyme complex consisting of ten subunits. investigations of the biochemical and structural properties of dna polymerase iii require the expression and purification of subunits including alpha, epsilon, theta, gamma, delta', delta, and beta separately followed by in vitro reconstitution of the pol iii core and clamp loader. here we propose a new method for expressing and purifying dna polymerase iii components by utilizing a protein co-expression strategy. our results show that the subunits of the pol iii core and those of the clamp loader can be co-expressed and purified based on inherent interactions between the subunits. the resulting pol iii core, clamp loader and sliding clamp can be reconstituted effectively to perform dna polymerization. our strategy considerably simplifies the expression and purification of dna polymerase iii and provides a feasible and convenient method for exploring other multi-subunit systems. |
| WOS关键词 | CHROMOSOMAL REPLICATION MACHINE ; ONE HOLOENZYME PARTICLE ; CLAMP LOADER ; SLIDING CLAMPS ; ESCHERICHIA-COLI ; TEMPERATURE ; COMPONENTS |
| WOS研究方向 | Cell Biology |
| WOS类目 | Cell Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000310519800007 |
| 出版者 | HIGHER EDUCATION PRESS |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2375946 |
| 专题 | 武汉病毒研究所 |
| 通讯作者 | Bi, Li-Jun |
| 作者单位 | 1.Chinese Acad Sci, Natl Lab Biomacromol, Inst Biophys, Beijing 100101, Peoples R China 2.Chinese Acad Sci, State Key Lab Virol, Wuhan Inst Virol, Wuhan 430071, Hubei Province, Peoples R China 3.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China |
| 推荐引用方式 GB/T 7714 | Lin, Shi-Qiang,Bi, Li-Jun,Zhang, Xian-En. A simplified method for reconstituting active e. coli dna polymerase iii[J]. Protein & cell,2011,2(4):303-307. |
| APA | Lin, Shi-Qiang,Bi, Li-Jun,&Zhang, Xian-En.(2011).A simplified method for reconstituting active e. coli dna polymerase iii.Protein & cell,2(4),303-307. |
| MLA | Lin, Shi-Qiang,et al."A simplified method for reconstituting active e. coli dna polymerase iii".Protein & cell 2.4(2011):303-307. |
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来源：武汉病毒研究所
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