High-level heterologous gene expression in Ochrobactrum anthropi using an A-rich UP element.

DNA regions that flank a gene's promoter play an important role in determining transcription efficiency by interacting with the carboxy-terminal domain of RNA polymerase alpha-subunit. We placed an adenine-rich upstream element (UP) between -38 and -59 of the core trc promoter to enhance gene expression in Ochrobactrum anthropi up to 66-fold. The high level of expression achieved by the UP element and the N-terminus fusion of a 6xHis epitope tag facilitated detection and purification of heterologous proteins directly from O. anthropi.