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.
Seleem
Mohamed
M
Department of Biomedical Sciences and Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, 1410 Prices Fork Rd, Blacksburg, VA 24061-0342, USA.
Ali
Mohammed
M
Abd Al-Azeem
M W
MW
Boyle
Stephen M
SM
Sriranganathan
Nammalwar
N
D015153:Blotting, Western; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D017930:Genes, Reporter; D008967:Molecular Biology / Q000379:methods; D008969:Molecular Sequence Data; D020621:Ochrobactrum anthropi / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D050436:Regulatory Elements, Transcriptional / Q000235:genetics* / Q000502:physiology; D017422:Sequence Analysis, DNA; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D001616:beta-Galactosidase