Distance between RBS and AUG plays an important role in overexpression of recombinant proteins.
The spacing between ribosome binding site (RBS) and AUG is crucial for efficient overexpression of genes when cloned in prokaryotic expression vectors. We undertook a brief study on the overexpression of genes cloned in Escherichia coli expression vectors, wherein the spacing between the RBS and the start codon was varied. SDS-PAGE and Western blot analysis indicated a high level of protein expression only in constructs where the spacing between RBS and AUG was approximately 40 nucleotides or more, despite the synthesis of the transcripts in the representative cases investigated.
Berwal
Sunil K
SK
Cell and Molecular Biology Laboratory, Department of Biotechnology, University of Pune, Pune 411007, India.
Sreejith
R K
RK
Pal
Jayanta K
JK
D001665:Binding Sites; D018387:Codon, Initiator; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism*
D018387:Codon, Initiator; D011994:Recombinant Proteins