The analysis of promoters and the effective expression of exogenous proteins in a cold-adapted bacterium / 生物工程学报

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.