Supplementary Material for: Relative Strengths and Regulation of Different Promoter-Associated Sequences for Various blaSHV Genes and Their Relationships to β-Lactam Resistance

Aims: This work investigated the relative strengths of different blaSHV promoter-associated sequences and their regulation function in blaSHV expression and β-lactam resistance. Methods: Recombinant plasmids with the promoter-associated sequences (P-W, P-S, P-IS, and P-WPD), tac promoter, and combined fragments of promoter and blaSHV were separately constructed and transformed into Escherichia coli
DH5α. The relative strengths of the promoters indicated by the
intensities of green fluorescent protein and the mRNA expression levels
of blaSHV were compared. The minimum inhibitory concentration and extended spectrum β-lactamase phenotypes were evaluated. Results: The relative strengths were ranked as P-tac
> P-WPD > P-IS > P-S > P-W. The mRNA expression and
β-lactam resistance levels of the different promoter-associated sequence
groups were generally consistent with the strength rank, but the extent
of gfp and blaSHV mRNA levels varied
significantly in each group. The β-lactam resistance levels were
inconsistent with the strength rank in certain blaSHV groups. In relation to the different promoter-associated sequences, blaSHV-ESBLs displayed significantly different change modes of β-lactam resistance compared with blaSHV-non-ESBLs. Conclusion: The mRNA expression and β-lactam resistance of the blaSHV
showed consistencies and inconsistencies with the strengths of the
promoter-associated sequences. The mechanisms accounting for these
discrepancies need further investigation.