| 학회 |
한국화학공학회 |
| 학술대회 |
2006년 가을 (10/27 ~ 10/28, 고려대학교) |
| 권호 |
12권 2호, p.1778 |
| 발표분야 |
생물화공 |
| 제목 |
Co-expression of recombinant proteins in cell-free protein synthesis reactions |
| 초록 |
In our experiments to produce different combinations of recombinant proteins in a cell-free protein synthesis system derived from E.coli, we found that certain pairs of the ORFs were not expressed equally. Instead, only a single DNA species was expressed dominantly while the expression of the other one was almost completely repressed. Such a bias during the co-expression of DNA pairs was eliminated when an identical downstream box sequence was added to the 5’-ends of the template DNA pairs. Using the identical nucleotide sequences of his-tag or the downstream box of chloramphenicol acetyltransferase (CAT-DB) in front of the target genes that had not been otherwise expressed compatibly, both of the encoded proteins were produced at similar productivities. Moreover, in the presence of a common downstream box, multiple numbers of genes were simultaneously expressed in a same reaction mixture. We expect that the proposed approach will offer a powerful tool for the preparation of unbiased protein libraries as well as for studying the structure and functions of interacting proteins. |
| 저자 |
금정원1, 김태완1, 오린석1, 안진호1, 최차용1, 김동명2
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| 소속 |
1서울대, 2충남대 |
| 키워드 |
cell-free protein synthesis; coexpression
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| E-Mail |
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| 원문파일 |
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