Large-scale production and characterization of Bacillus thuringiensis subsp. tenebrionis insecticidal protein from Escherichia coli |
| |
| Authors: | M E Gustafson R A Clayton P B Lavrik G V Johnson R M Leimgruber S R Sims D E Bartnicki |
| |
| Institution: | (1) Monsanto Company, Searle Research and Development, 700 Chesterfield Parkway North, St. Louis, MO 63198, USA. Fax: +314537 6198, US;(2) Monsanto Company, Monsanto Ceregen Division, 700 Chesterfield Parkway North, St. Louis, MO 63198, USA, US;(3) Monsanto Company, Monsanto Corporate Research, 700 Chesterfield Parkway North, St. Louis, MO 63198, USA, US |
| |
| Abstract: | Bacillus thuringiensis subsp. tenebrionis insecticidal protein was produced in recombinant Escherichia coli and purified to near homogeneity to provide quantities of protein for safety-assessment studies associated with the registration
of transgenic potato plants. The 68-kDa protein is produced naturally by Bacillus thuringiensis subsp. tenebrionis by translation initiation at an internal initiation site in the native DNA sequence. The gene sequence specific for this
truncated protein was expressed in E. coli strain JM 101 and fermented at the 1000-l scale. The protein accumulated as insoluble inclusion bodies, and was purified
by extraction at pH␣10.8 with carbonate buffer, selective precipitation at pH 9.0, and differential centrifugation. No chromatography
steps were required to produce over 50 g purified protein as a lyophilized powder with a purity greater than 95 % and demonstrating
full insecticidal activity against Colorado potato beetle larvae. The protein was further characterized to assure identity
and suitability for use in safety-assessment studies.
Received: 31 May 1996 / Received revision: 11 September 1996 / Accepted: 13 October 1996 |
| |
| Keywords: | |
| 本文献已被 SpringerLink 等数据库收录！ |
|
