High-level heterologous gene expression in Ochrobactrum anthropi using an A-rich UP element
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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 α-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.
KeywordsMethylparathion Miller Unit Brucella Abortus Upstream Element Express Fusion Protein
We express our appreciation to Dr. Tim Larson of Virginia Tech for his critical reading of the manuscript and suggestions. We would like to thank Mrs. Kay Carlson of Virginia tech for providing us with O. anthropi strain and the support throughout the experiment.
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