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Special (비정기세미나)
2004.04.02 08:50

제6회 정기세미나

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Extra Form
1. 연사 : Prof. Yaeta Endo (Ehime University)
2. 연제 : High-Throughput, Gemone-Scale Protein Production
Method Based on the Wheat Germ Cell-Free Expression System
3. 일시 : 2004년 4월 6일 (화) 오후 4:00
4. 장소 : 의과학센터 원격강의실

※ 문의 : 유욱준 교수님 (2626)

※ Abstract
Current cell-free protein synthesis systems can synthesize
proteins with high speed and accuracy, but produce only a low yield
because of their instability over time. Here I describe the protein
synthesis method with a highly efficient but also robust cell-free
system from wheat embryos. We first investigated the source of the
instability of existing systems in light of endogenous
ribosome-inactivating proteins. We found that conventional wheat germ
extracts contained the RNA N-glycosidase tritin and other inhibitors
such as thionin, ribonucleases, deoxyribonucleases, and proteases, that
originate from the endosperm inhibit translation. Extensive washing of
wheat embryos, to eliminate endosperm contaminants, has resulted in
extracts with a high degree of stability and activity. In order to
maximize the translation yield and throughput of the system, we
addressed and resolved the following issues: optimization of the ORF
flanking regions, a new strategy to construct PCR-generated DNAs for
screening, and design of an expression vector for large-scale protein
production. One possible procedure for the preparation of proteins is
as follows: (1) selection of suitable genes from a protein catalogue
made by the PCR-directed system; (2) cloning into pEU and transcription
of mRNA; and finally (3) protein synthesis in system.
The cell-free system bypasses living processes that has made us
possible to invent two robotic systems, GenDecoder and Protemist for
HTP-screening and for massive preparation of protein. We will be
discussing many possibilities and avenues in the post-genome era with
respect to our technology.

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