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Translation of vph mRNA in Streptomyces lividans and Escherichia coli after removal of the 5′ untranslated leader
Author(s) -
Wu ChiJu,
Janssen Gary R.
Publication year - 1996
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1046/j.1365-2958.1996.00119.x
Subject(s) - biology , start codon , shine dalgarno sequence , open reading frame , ribosomal binding site , transcription (linguistics) , eukaryotic translation , untranslated region , coding region , stop codon , five prime untranslated region , genetics , microbiology and biotechnology , messenger rna , gene , translation (biology) , peptide sequence , philosophy , linguistics
The Streptomyces vinaceus viomycin phosphotransferase ( vph ) mRNA contains an untranslated leader with a conventional Shine–Dalgarno homology. The vph leader was removed by ligation of the vph coding sequence to the transcriptional start site of a Strepto‐myces or an Escherichia coli promoter, such that transcription would initiate at the first position of the vph start codon. Analysis of mRNA demonstrated that transcription initiated primarily at the A of the vph AUG translational start codon in both Streptomyces lividans and E. coli ; cells expressing the unleadered vph mRNA were resistant to viomycin indicating that the Shine–Dalgarno sequence, or other features contained within the leader, was not necessary for vph translation. Addition of four nucleotides (5′‐AUGC‐3′) onto the 5′ end of the unleadered vph mRNA resulted in translation initiation from the vph start codon and the AUG triplet contained within the added sequence. Translational fusions of vph sequence to a Tn 5 neo reporter gene indicated that the first 16 codons of vph coding sequence were sufficient to specify the translational start site and reading frame for expression of neomycin resistance in both E. coli and S. lividans.