Effect of a rare leucine codon, TTA, on expression of a foreign gene in Streptomyces lividans.
Streptomyces are bacteria with a very high chromosomal G+C composition (> 70 mol%) and extremely biased codon usage. In order to investigate the relationship between codon usage and gene expression in Streptomyces, we used ssi (Streptomyces subtilisin inhibitor) as a reporter gene and monitored its secretory expression in S. lividans. In consequence of alteration of the native codons of Leu, Lys and Ser of ssi to minor ones by site-directed mutagenesis, i.e., Leu79-Leu80: CTG-CTC to TTA-TTA, Lys89: AAG to AAA, Ser108-Ser109: TCG-AGC to TCT-TCT, respectively, the production of SSI was reduced remarkably in the case of TTA codons, while it was slightly increased in the case of AAA and almost the same in TCT codons. This conspicuous decrease found for Leu codon replacement was probably due to the low availability of intracellular tRNA(Leu) (UUA), a product of bldA which has been reported to be expressed only during the late stage of growth.
Ueda
Y
Y
Department of Industrial Chemistry, Faculty of Engineering, University of Tokyo, Japan.
Taguchi
S
S
Nishiyama
K
K
Kumagai
I
I
Miura
K
K
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D003062:Codon; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D007930:Leucine / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D013302:Streptomyces / Q000235:genetics*; D013381:Subtilisins / Q000037:antagonists & inhibitors*; D014162:Transfection
D001426:Bacterial Proteins; D003062:Codon; C018150:subtilisin inhibitor protein, Streptomyces; D013381:Subtilisins; D007930:Leucine