[The analysis of promoters and the effective expression of exogenous proteins in a cold-adapted bacterium].
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.
Wei
Yunlin
Y
Biotechnology Research Center of Kunming University of Science and Technology, Kunming 650224, China. weiyunlin@yahoo.com.cn
Ji
Xiuling
X
Lin
Lianbing
L
Jing
Shenrong
S
D000150:Acinetobacter / Q000235:genetics* / Q000378:metabolism; D000222:Adaptation, Physiological / Q000235:genetics*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D003080:Cold Temperature; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014170:Transformation, Genetic; D000516:alpha-Amylases / Q000096:biosynthesis* / Q000235:genetics
D001426:Bacterial Proteins; D000516:alpha-Amylases