Large-scale production of soluble recombinant amyloid-beta peptide 1-42 using cold-inducible expression system
- Authors
- Kim, Eun-Kyung; Moon, Jeong Chan; Lee, Jeong Mi; Jeong, Min Seop; Oh, Choongseob; Ahn, Sung-Min; Yoo, Yung Joon; Jang, Ho Hee
- Issue Date
- Nov-2012
- Publisher
- ACADEMIC PRESS INC ELSEVIER SCIENCE
- Keywords
- Aggregation-prone; Alzheimer' s disease; Amyloid-beta peptide 1-42; Cell viability; Cold induction; Soluble expression
- Citation
- PROTEIN EXPRESSION AND PURIFICATION, v.86, no.1, pp.53 - 57
- Journal Title
- PROTEIN EXPRESSION AND PURIFICATION
- Volume
- 86
- Number
- 1
- Start Page
- 53
- End Page
- 57
- URI
- https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/16046
- DOI
- 10.1016/j.pep.2012.08.021
- ISSN
- 1046-5928
- Abstract
- Amyloid-beta peptide 1-42 (A beta(1-42)), the predominant form in senile plaques, plays important roles in the pathogenesis of Alzheimer's disease. Because A beta(1-42) has aggregation-prone nature, it has been difficult to produce in a soluble state in bacterial expression systems. In this study, we modified our expression system to increase the soluble fraction of A beta(1-42) in Escherichia coli (E. coli) cells. The expression level and solubility of recombinant A beta(1-42) induced at the low temperature (16 degrees C) is highly increased compared to that induced at 37 degrees C. To optimize expression temperature, the coding region of A beta(1-42) was constructed in a pCold vector, pCold-TF, which has a hexahistidine-tagged trigger factor (TF). Recombinant A beta(1-42) was expressed primarily as a soluble protein using pCold vector system and purified with a nickel-chelating resin. When the toxic effect of recombinant A beta(1-42) examined on human neuroblastoma SH-SY5Y cells, the purified A beta(1-42) induced cell toxicity on SH-SY5Y cells. In conclusion, the system developed in this study will provide a useful method for the production of aggregation prone-peptide such as A beta(1-42). (C) 2012 Elsevier Inc. All rights reserved.
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