电邮这篇文章 Optimization of rhBMP-2 active-form production in a heterologous expression system using microbiological and molecular genetic approaches
- 作者: Karyagina A.S.1,2,3, Boksha I.S.1, Grunina T.M.1, Demidenko A.V.1, Poponova M.S.1, Sergienko O.V.1,3, Lyaschuk A.M.1, Galushkina Z.M.1, Soboleva L.A.1, Osidak E.O.1,4, Semikhin A.S.1, Gromov A.V.1, Lunin V.G.1,3
-
隶属关系:
- Gamaleya Federal Research Center for Epidemiology and Microbiology
- Belozersky Research Institute of Physico-Chemical Biology
- All-Russia Research Institute of Agricultural Biotechnology
- Imtek Ltd.
- 期: 卷 31, 编号 4 (2016)
- 页面: 208-213
- 栏目: Experimental Works
- URL: https://journals.rcsi.science/0891-4168/article/view/178130
- DOI: https://doi.org/10.3103/S0891416816040030
- ID: 178130
如何引用文章
开放存取
##reader.subscriptionAccessGranted##
订阅存取
详细
Recombinant bone morphogenetic protein-2 (rhBMP-2) has pronounced osteoinductive properties, as evidenced by the results of experimental and clinical practices. This applies to both the protein produced in eukaryotic cells and the protein synthesized in bacterial cells. In eukaryotic expression systems, production of the protein is extremely low and, consequently, the cost of materials on its basis is very high. Therefore, optimization of heterologous expression systems for rhBMP-2 production represents an important task. In the present work, optimization of codon composition of the rhBMP-2 gene nucleotide sequence and secondary structure of the transcript, as well as strain selection for efficient gene expression, were carried out. The producing strain based on Escherichia coli BL-21(DE3) provides a high level of rhBMP-2 synthesis (about 57% of total cell proteins). Biological activity of rhBMP-2 dimeric forms purified from the obtained producing strain was measured by induction of alkaline phosphatase activity in C2C12 cells. It is comparable with that of commercial rhBMP-2 expressed in E. coli (R&D Systems, United States). Purified rhBMP-2 does not contain impurities of E. coli endotoxin and can be used in experimental studies of osteoinduction in laboratory animals.
作者简介
A. Karyagina
Gamaleya Federal Research Center for Epidemiology and Microbiology; Belozersky Research Institute of Physico-Chemical Biology; All-Russia Research Institute of Agricultural Biotechnology
编辑信件的主要联系方式.
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098; Moscow, 119991; Moscow, 127550
I. Boksha
Gamaleya Federal Research Center for Epidemiology and Microbiology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098
T. Grunina
Gamaleya Federal Research Center for Epidemiology and Microbiology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098
A. Demidenko
Gamaleya Federal Research Center for Epidemiology and Microbiology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098
M. Poponova
Gamaleya Federal Research Center for Epidemiology and Microbiology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098
O. Sergienko
Gamaleya Federal Research Center for Epidemiology and Microbiology; All-Russia Research Institute of Agricultural Biotechnology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098; Moscow, 127550
A. Lyaschuk
Gamaleya Federal Research Center for Epidemiology and Microbiology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098
Z. Galushkina
Gamaleya Federal Research Center for Epidemiology and Microbiology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098
L. Soboleva
Gamaleya Federal Research Center for Epidemiology and Microbiology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098
E. Osidak
Gamaleya Federal Research Center for Epidemiology and Microbiology; Imtek Ltd.
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098; Moscow, 121552
A. Semikhin
Gamaleya Federal Research Center for Epidemiology and Microbiology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098
A. Gromov
Gamaleya Federal Research Center for Epidemiology and Microbiology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098
V. Lunin
Gamaleya Federal Research Center for Epidemiology and Microbiology; All-Russia Research Institute of Agricultural Biotechnology
Email: akaryagina@gmail.com
俄罗斯联邦, Moscow, 123098; Moscow, 127550
补充文件
