지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2013.1
- 수록면
- 169 - 177 (9page)
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Recently, the number of people with diabetes is rapidly increasing, coupled with the fact that the insulin market is remarkably increasing. Therefore, molecular farming for plant-derived pharmaceutical protein production is reported as becoming more attractive than ever. In this study, we carried out experiments step by step for development of recombinant insulin constructs, which were transformed into E. coli system, in vitro transcription and translation system, and tobacco cells. At first, recombinant proinsulin protein was successfully produced in in vitro transcription and translation system with wheat germ extract. After which, recombinant construct of prominiinsulin encoded a fusion protein of 7.8 kDa with trypsin cleavage sites at N terminus and C terminus of minimized C-peptide was tried to in vitro expression using E.coli culture. After purification with His-tag column, the resulting recombinant prominiinsulin protein was processed with trypsin, and then checked insulin biosynthesis by SDS-PAGE and western blot analysis with anti-insulin monoclonal antibody. The immunoreactive product of trypsin-treated miniinsulin was identical to the predicted insulin hexamer. The construct of 35S promoter-driven preprominiinsulin recombinant gene with signal peptide region for ER-targeting and red fluorescence protein gene [N terminus → tobacco E2 signal peptide → B-peptide (1-29 AA) → AAK → A-peptide (1-21 AA) → RR → His6 → KDEL → C terminus] was transformed into BY-2 tobacco cells. A polypeptide corresponding to the 38-kDa molecular mass predicted for fusion protein was detected in total protein profiles from transgenic BY-2 cells by western analysis. Therefore, this recombinant preprominiinsulin construct can be used for generation of transgenic tobacco plants producing therapeutic recombinant insulin.
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참고문헌 (20)
참고문헌 신청
/ 2006 / WO 10-2006-0052705 (2006) Methods for the production of insulin in plants. SemBioSys Genetics Incorporated
Cardinale F / 2002 / Convergence and Divergence of Stress-Induced Mitogen-Activated Protein Kinase Signaling Pathways at the Level of Two Distinct Mitogen-Activated Protein K
Cornelissen BJ / 1986 / Bol JF(1986)A tobacco mosaic virus-induced tobacco protein is homologous to the sweet-tasting protein thaumatin / Nature 321 : 531 ~ 532
Cowley DJ / 1997 / Expression, purification and characterization of recombinant human proinsulin / FEBS Lett 402 : 124 ~ 130
Dunn MF / 2005 / Zinc-ligand interactions modulate assembly and stability of the insulin hexamer - a review / Biometals 18 : 295 ~ 303
Cardinale F / 2002 / Convergence and Divergence of Stress-Induced Mitogen-Activated Protein Kinase Signaling Pathways at the Level of Two Distinct Mitogen-Activated Protein K
Cornelissen BJ / 1986 / Bol JF(1986)A tobacco mosaic virus-induced tobacco protein is homologous to the sweet-tasting protein thaumatin / Nature 321 : 531 ~ 532
Cowley DJ / 1997 / Expression, purification and characterization of recombinant human proinsulin / FEBS Lett 402 : 124 ~ 130
Dunn MF / 2005 / Zinc-ligand interactions modulate assembly and stability of the insulin hexamer - a review / Biometals 18 : 295 ~ 303
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