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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
- 2021.1
- 수록면
- 309 - 316 (8page)
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초록· 키워드
오류제보하기
Plants are one of the ideal models for therapeutic protein production, however the recombinant protein purifcation problems in them must be overcome. Bone Morphogenetic Protein2 (BMP2) is employed for the restoration and construction of bone tissues. Hydrophobin is a fungal based protein with high hydrophobic characteristics. Due to this specifcity, it is suitable for the purifcation of chimer protein from complex solutions when is fused to a protein utilizing an aqueous two-phase (A2P) technique. The plant optimized mature human BMP2 gene was designed and evaluated by in silico method. This process involves simulating molecular dynamics using the RMSD, RMSF and Gyration radius indexes. The synthesized Hyd-BMP2 gene was cloned into a pTRAkc-ERH plasmid and Transferred into Agrobacterium (Gv3101). The Nicotiana benthamiana plant leaves were co-agroinfltrated with HA-Hyd-BMP2 and P19-pCambia1304 containing silencing suppressor. After purif?cation of plant extract utilizing the A2P method, the sample was subjected to SDS-PAGE and Western-blot. By in silico study, the simulated fusion protein proftably shows reasonable protein compactness and the efect of amino acid substitution on protein?protein interaction is not remarkable. Western-blotting using anti HA tag has shown that the A2P technique partially purifed the two 22?kDa and 44?kDa forms of Hydrophobin-BMP2. These results confrmed the presence of monomer and dimer forms of Hydrophobin-BMP2 proteins. Moreover, the expression level of the protein using P19 silencing suppressor increased six times and to 0.018% as shown by ELISA. This study presents a fast and easy technique for the purifcation of transient expressed pharmaceutical proteins from plants.
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참고문헌 (47)
참고문헌 신청
Bedrov D / 2019 / Molecular dynamics simulations of ionic liquids and electrolytes using polarizable force fields / Chem Rev 119:7940~7995
Berger BW / 2019 / Hydrophobins: multifunctional biosurfactants for interface engineering / J Biol Eng 13:10
Bhargava BL / 2005 / Dynamics in a room-temperature ionic liquid: a computer simulation study of 1,3-dimethylimidazolium chloride / J Chem Phys 123:144505
Chen H / 1994 / Enhanced recovery of transformants of Agrobacterium tumefaciens after freeze-thaw transformation and drug selection / Biotechniques 16(664?8):70
Childers MC / 2018 / Validating molecular dynamics simulations against experimental observables in light of underlying conformational ensembles / J Phys Chem B 122:6673~6689
Berger BW / 2019 / Hydrophobins: multifunctional biosurfactants for interface engineering / J Biol Eng 13:10
Bhargava BL / 2005 / Dynamics in a room-temperature ionic liquid: a computer simulation study of 1,3-dimethylimidazolium chloride / J Chem Phys 123:144505
Chen H / 1994 / Enhanced recovery of transformants of Agrobacterium tumefaciens after freeze-thaw transformation and drug selection / Biotechniques 16(664?8):70
Childers MC / 2018 / Validating molecular dynamics simulations against experimental observables in light of underlying conformational ensembles / J Phys Chem B 122:6673~6689
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