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. 2019 Jun 26;10:1454. doi: 10.3389/fmicb.2019.01454

FIGURE 1.

FIGURE 1

The strategy used for constructing a series of lac promoter and ara promoter reporter vectors. (A) Schematic representation of wild-type β-galactosidase, lacZM15, and lacZα polypeptides. Five distinct domains of the β-galactosidase monomer (1024 aa) was marked. LacZM15 is a deletion mutant (residues 11–41) that maps in the α region. The preferred α-donor peptide lacZα involved in this study contains wild type residues 1–56. α-complemented active β-galactosidase assembled from lacZM15 and lacZα contains two sets of overlapping sequences, which are segments 1–10 and 42–56. (B) Assembly strategy for the lac promoter and ara promoter reporter vectors used in the study. Assembly of the desired target promoter, ribosome binding site, a reporter gene (including the encoding sequence for lacZα, mCherry, or full-length lacZ), and rrnB terminator via the genetic methods to construct the reporter cassette. The resulting fusion fragments are then inserted into pBR322 via NdeI and EcoRI sites.