| 학회 | 한국화학공학회 |
| 학술대회 | 2005년 가을 (10/21 ~ 10/22, 인하대학교) |
| 권호 | 11권 2호, p.2029 |
| 발표분야 | 생물화공 |
| 제목 | Rapid detection method for protein-protein interaction in a cell-free protein synthesis system |
| 초록 | We have developed a simple method for the analysis of protein-protein interaction based on a highly efficient cell-free protein synthesis system. In this method, genes of potential prey proteins are co-expressed with a affinity-tagged bait protein in a cell-free protein synthesis reaction. In addition, synthesis reactions were conducted in the presence of affinity beads so that the bait-prey complex can be in situ isolated onto the beads. As a model system to validate this approach, human EPO and EPO receptor (histidine-tagged) were cell-free synthesized simultaneously in the presence of Ni-NTA magnetic agarose beads. Mass spectrometry and immunoblotting analysis of the eluates confirmed the presence of EPO and EPO-receptor in the eluates from the beads indicating the successful isolation of the interacting proteins. We expect the developed method will be easily expanded for parallel analysis of protein interactions. |
| 저자 | 김태완1, 오인석1, 금정원1, 최차용1, 김동명2 |
| 소속 | 1서울대, 2충남대 |
| 키워드 | cell-free protein synthesis |
| 원문파일 | 초록 보기 |
