Codon optimization of the gene encoding a domain from human type 1 neurofibromin protein results in a threefold improvement in expression level in Escherichia coli

An internal domain from the human type 1 neurofibromin has previously been expressed in Escherichia coli as a fusion with gluthathione S-transferase (GST). The expression level of this protein was lower than expected and so a gene was constructed using the distribution of codons found in highly expr...

Full description

Saved in:
Bibliographic Details
Published in:Protein expression and purification 1998-03, Vol.12 (2), p.185-188
Main Authors: Hale, R S, Thompson, G
Format: Article
Language:English
Subjects:
Base Sequence
Codon - chemistry
Codon - genetics
Electrophoresis, Polyacrylamide Gel
Escherichia coli - genetics
Gene Expression Regulation - genetics
Humans
Molecular Sequence Data
Neurofibromin 1
Protein Biosynthesis
Proteins - chemistry
Proteins - genetics
Recombinant Proteins - biosynthesis
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Sequence Alignment
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:An internal domain from the human type 1 neurofibromin has previously been expressed in Escherichia coli as a fusion with gluthathione S-transferase (GST). The expression level of this protein was lower than expected and so a gene was constructed using the distribution of codons found in highly expressed E. coli proteins. Codons were assigned using a Microsoft Visual Basic computer program to give a distribution similar to those found in genes which are highly expressed in E. coli. The optimized gene was then cloned back into the same GST fusion plasmid and it was found that the expression of soluble protein had increased threefold.
ISSN:1046-5928
DOI:10.1006/prep.1997.0825