Gene expression signals derived from Lactococcus lactis were linked to lacZ‐fused genes with different 5′‐nucleotide sequences. Computer predictions of mRNA secondary structure were combined with lacZ expression studies to direct base‐substitutions that could possibly influence gene expression. Mutations were made such that the DNA sequence upstream of the ATG start codon was not changed. Moreover, care was taken that the substitutions, which were all within the first six codons, neither affected the amino acid sequence of the gene product nor introduced codons rarely used in L. lactis. The results suggest that mRNA secondary structure contributes to the efficiency of translation initiation in L. lactis.
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