Codon adjustment to maximise heterologous gene expression in Streptomyces lividans can lead to decreased mRNA stability and protein yield.
The impact of the codon bias of the mouse tumour necrosis factor alpha (mTNF) gene cloned in Streptomyces lividans on the efficiency of expression and secretion was analysed. Minor codons occurring in the mTNF gene were therefore adapted to the codon bias of Streptomyces by site-directed mutagenesis. No improvement in mTNF yield could be detected. The stability of the transcript derived from the construct was shown to be more important for determining the final level of mTNF production. A strong correlation was observed between the yield of secreted biologically active mTNF and the amount of mTNF mRNA present in the cells.
Lammertyn
E
E
Rega Institute, Katholieke Universiteit Leuven, Belgium.
Van Mellaert
L
L
Bijnens
A P
AP
Joris
B
B
Anné
J
J
D000465:Algorithms; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics*; D018387:Codon, Initiator / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D011401:Promoter Regions, Genetic / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012984:Software; D013302:Streptomyces / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism
D003062:Codon; D018387:Codon, Initiator; D017931:DNA Primers; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D000516:alpha-Amylases