A. Saraffova et al., COMPARATIVE-STUDY ON THE EFFECT OF SIGNAL PEPTIDE CODONS AND ARGININECODONS ON THE EXPRESSION OF HUMAN INTERFERON-ALPHA-1 GENE IN ESCHERICHIA-COLI, Journal of interferon & cytokine research, 16(9), 1996, pp. 745-749
Human interferon-alpha 1 (HuIFN-alpha 1) gene containing signal peptid
e codons is poorly expressed in bacteria, and this is explained by the
presence of clusters of rare (AGG) arginine codons in its structure,
In this study, we have constructed a series of modified HuIFN-alpha 1
genes to study the effect of both residual signal peptide codons and c
lusters of AGG codons on gene expression in Escherichia coil cells, Ou
r results showed that substitution of preferential for rare arginine c
odons in two clusters did not affect the yield, whereas deletion of th
e signal peptide codons led to a 10-fold increase in the yield of reco
mbinant protein, To understand the mechanism of interference of gene s
tructure on the expression of the HuIFN-alpha 1 gene in vivo, both the
level and stability of HuIFN-alpha 1 mRNA were measured, The amount o
f HuIFN mRNA increased almost five times on deletion of the signal pep
tide codons from HuIFN-alpha 1 gene constructs (containing AGG cluster
s or not), The stability of mRNA obtained from all gene constructs was
shown to be the same (half-life of 60 +/- 5 sees), indicating that th
e signal peptide codons interfere with both the efficiency of transcri
ption of the HuIFN-alpha 1 gene and translation of its mRNA.