Group II Intron Protein Localization and Insertion Sites Are Affected by Polyphosphate (original) (raw)
Figure 1
Intron-Expression Plasmid and Cell Microarrays Used to Identify E. coli Disruptants with Altered GFP/LtrA Localization
**(**A) pACD2X-GFP/LtrA uses a T7 lac promoter to express the Ll.LtrB-ΔORF intron with short flanking exons, followed by a GFP/LtrA fusion protein [12]. E1 and E2 are 5′ and 3′ exons, respectively, and T1 and T2 are rrnB transcription terminators.
(B) Wide-field light scattering image of a cell microarray. A mariner transposon-insertion library of E. coli HMS174(DE3) cells carrying pACD2X-GFP/LtrA was arrayed onto microscope slides like that shown and screened by automated fluorescence microscopy to identify mutants with altered GFP/LtrA localization patterns (see Materials and Methods section).
(C) Close-up of a spot from the cell microarray.
(D) Higher magnification view of the same spot focusing on an E. coli cell with the wild-type bipolar GFP/LtrA localization pattern.