Codon usage as a reason for unsuccessful search for amber-suppressor mutants inStreptomyces lividans?

Abstract

SummaryAn amber mutation was created in the CAT gene of plasmid vector pACYC184 and this modified plasmid was fused with theStreptomycesvector pIJ702 for use as an indicator for the identification ofStreptomycesstrains carrying nonsense suppressor tRNA mutations. The resulting hybrid plasmid pGM1109 was introduced into the chloramphenicol-sensitive mutant M252 ofStreptomyces lividans. Chloramphenicol-resistant colonies were isolated and characterized. None of them was a nonsense suppressor mutant. The failure to obtain such mutants is discussed on the basis of codon usage in streptomycetes.