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A new cloning strategy for generating multiple repeats of a repetitive polypeptide based on non-template PCR

Authors
Chu, Hun-SuRyum, JungPark, Seo-YoungKim, Byung-GeeKim, Dong-MyungWon, Jong-In
Issue Date
May-2011
Publisher
SPRINGER
Keywords
Elastin-like polypeptide (ELP); Length library; Multiple repeats of amino acids; Non-template PCR
Citation
BIOTECHNOLOGY LETTERS, v.33, no.5, pp.977 - 983
Journal Title
BIOTECHNOLOGY LETTERS
Volume
33
Number
5
Start Page
977
End Page
983
URI
https://scholarworks.bwise.kr/hongik/handle/2020.sw.hongik/19887
DOI
10.1007/s10529-010-0510-7
ISSN
0141-5492
Abstract
A new cloning method for generating multiple repeats of amino acids is described which can be used as biomaterials, protein polymers and biomedical applications. Although several traditional methods for cloning multiple repeats are still exploited, these are laborious and complicated because they must go through several consecutive cloning steps. To solve these problems, synthetic gene libraries encoding repetitive patterns were constructed by using non-template PCR. As a result, a 'length library' with fourteen different ELP repeating genes was constructed and expressed in a cell-free protein synthesis system. These results showed our novel cloning method is efficient, and has the potential capacity for synthesizing repetitive genes by PCR to be cloned in any commercial expression vectors.
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