The analysis of promoters and the effective expression of exogenous proteins in a cold-adapted bacterium.
- Author:
Yunlin WEI
1
;
Xiuling JI
;
Lianbing LIN
;
Shenrong JING
Author Information
1. Biotechnology Research Center of Kunming University of Science and Technology, Kunming 650224, China. weiyunlin@yahoo.com.cn
- Publication Type:Journal Article
- MeSH:
Acinetobacter;
genetics;
metabolism;
Adaptation, Physiological;
genetics;
Bacterial Proteins;
biosynthesis;
genetics;
Base Sequence;
Cold Temperature;
Escherichia coli;
genetics;
metabolism;
Genetic Vectors;
genetics;
Molecular Sequence Data;
Promoter Regions, Genetic;
genetics;
Transformation, Genetic;
alpha-Amylases;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2008;24(3):415-422
- CountryChina
- Language:Chinese
-
Abstract:
Based on the constructed promoter probe vectors that could replicate both in E. coli and in a cold-adapted bacterium, several candidate promoters were isolated and their activities were evaluated by RT-PCR. The transcription initiation sites and core sequence of promoters were determined by primer extension analysis. A low-temperature expression vector was constructed by using the strongest promoter and a thermolabile alpha-amylase gene was successfully overproduced under control of this promoter at low temperature (7 degrees C), while the secreted alpha-amylase amounted up to 35% of the total extracellular proteins. The expression system is expected to be useful for the production of thermolabile exogenous proteins at low temperatures.
