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Comparative Study
. 1999 Oct;181(20):6569-72.
doi: 10.1128/JB.181.20.6569-6572.1999.

Efficiency of transcription from promoter sequence variants in Lactobacillus is both strain and context dependent

A McCracken  1 , P Timms
Affiliations
Comparative Study

Efficiency of transcription from promoter sequence variants in Lactobacillus is both strain and context dependent

A McCracken et al. J Bacteriol. 1999 Oct.

Abstract

The introduction of consensus -35 (TTGACA) and -10 (TATAAT) hexamers and a TG motif into the Lactobacillus acidophilus ATCC 4356 wild-type slpA promoter resulted in significant improvements (4.3-, 4.1-, and 10.7-fold, respectively) in transcriptional activity in Lactobacillus fermentum BR11. In contrast, the same changes resulted in decreased transcription in Lactobacillus rhamnosus GG. The TG motif was shown to be important in the context of weak -35 and -10 hexamers (L. fermentum BR11) or a consensus -10 hexamer (L. rhamnosus GG). Thus, both strain- and context-dependent effects are critical factors influencing transcription in Lactobacillus.

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Figures

FIG. 1
FIG. 1
Primer extension analysis of lactobacilli harboring the wild-type slpA promoter. Lane 1, L. rhamnosus GG; lane 2, L. fermentum BR11; lanes C, T, A, and G, sequencing reactions. Transcription was initiated from the start site published for the natural host L. acidophilus ATCC 4356 (2), with 4.5-fold-higher efficiency in L. rhamnosus GG than in L. fermentum BR11.
FIG. 2
FIG. 2
Northern blot analysis of RNA isolated from L. fermentum BR11 (A) and L. rhamnosus GG (B) harboring the slpA wild-type and derivative promoter constructs. Membranes were hybridized with DIG-labelled gusA and 16S rRNA probes as described in Materials and Methods. Lanes 1, pNZslp (wild type); lanes 2, pNZTG; lanes 3, pNZT10; lanes 4, pNZT35; lanes 5, pNZT1035; lanes 6, pNZS10; lanes 7, pNZS35; lanes 8, pNZS1035. Signals were quantified by densitometry and normalized for relative plasmid copy number. The relative transcriptional activities determined are given in Table 1.
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