Skip to main content
. 2019 Jun 5;10:1218. doi: 10.3389/fmicb.2019.01218

Figure 1.

Figure 1

(A) vgrG2b island organization. The genes are labeled with the given name (i.e., hcp2b) and are indicated by their annotation number (e.g., PA0263). The Sec signal peptide of Tli3 (PA0261) and the two DUF2875 of Tla3 (PA0259) are represented with stripped boxes. (B) The Tle3 periplasmic toxicity is counteracted by Tli3 (PA0261). Serial dilutions (from non-diluted to 10−7) of normalized cultures of E. coli BL21(DE3)pLysS producing the wild-type Tle3 in the cytoplasm, called Tle3c (from pVT1, a pETDuet-1 derivative) or in the periplasm, called Tle3p [from pSBC81, a pET22b(+) derivative yielding a fusion of Tle3 with a Sec signal peptide] were spotted on LB agar plates supplemented (left panel) with 0.4% glucose or (right panel) with 0.1 mM IPTG. Glucose and IPTG allow respectively repression and induction of the gene encoding the T7 RNA polymerase. When indicated Tli3 (PA0261) or Tla3 (PA0259) were produced in the periplasm from pVT8, pSBC107 and pVT9, respectively, pRSFDuet-1 derivatives. Line 1: pET22b(+) and pRSFDuet-1, line 2: pVT1 and pRSFDuet-1, line 3: pSBC81 and pRSFDuet-1, line 4: pSBC81 and pVT8, line 5: pSBC81 and pVT9, line 6: pVT8 and pET22b(+), line 7: pVT9 and pET22b(+), and line 8: pSBC107 is a pRSFDuet-1 derivative producing Tli3 (PA0261) and Tle3p from the same transcript.