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A plasmid toolbox for the easy autodisplay of recombinant proteins and its optimization.

Furtmann C, Rohe P, Gesing K, Kuss H, Lenz F, Jose J
Commun Biol. 2026 May 21;9(1):694. doi: 10.1038/s42003-026-10324-7. (Link opens in a new window) PubMed (Link opens in a new window) Article

The Autodisplay Toolbox (ATB) is a collection of plasmids (pATB) that enable the display of recombinant proteins at the surface of Gram-negative bacterial cells (autodisplay). Based on a classical (EhaA) and an inverse (YeeJ) autotransporter protein, variants were constructed with different signal peptides, linkers, and adjacent regions. To account for different expression needs, gene expression was set under control of an araBAD, a rhaBAD, or a pRox306 promoter resulting in 81 pATB in total. The ATB provides a simple option for optimization and can be used by experts but also opens up the world of bacterial surface display for non-experts.

Plasmids from Article

ID Plasmid Purpose
226014pATB_111Plasmid encodes an AT-FP with CtxB signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226015pATB_112Plasmid encodes an AT-FP with CtxB signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226016pATB_113Plasmid encodes an AT-FP with CtxB signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226017pATB_114Plasmid encodes an AT-FP with CtxB signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226018pATB_115Plasmid encodes an AT-FP with CtxB signal peptide, Δepitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226019pATB_116Plasmid encodes an AT-FP with CtxB signal peptide, ∆epitope∆β1∆-connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226020pATB_121Plasmid encodes an AT-FP with OprF signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226021pATB_122Plasmid encodes an AT-FP with OprF signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226022pATB_123Plasmid encodes an AT-FP with OprF signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226023pATB_124Plasmid encodes an AT-FP with OprF signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226024pATB_125Plasmid encodes an AT-FP with OprF signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226025pATB_126Plasmid encodes an AT-FP with OprF signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226026pATB_131Plasmid encodes an AT-FP with CtxB-spacer signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226027pATB_132Plasmid encodes an AT-FP with CtxB-spacer signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226028pATB_133Plasmid encodes an AT-FP with CtxB-spacer signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226029pATB_134Plasmid encodes an AT-FP with CtxB-spacer signal peptide, Delta-epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226030pATB_135Plasmid encodes an AT-FP with CtxB-spacer signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226031pATB_136Plasmid encodes an AT-FP with CtxB-spacer signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226032pATB_141Plasmid encodes an AT-FP with OprF-spacer signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226033pATB_142Plasmid encodes an AT-FP with OprF-spacer signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226034pATB_143Plasmid encodes an AT-FP with OprF-spacer signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226035pATB_144Plasmid encodes an AT-FP with OprF-spacer signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226036pATB_145Plasmid encodes an AT-FP with OprF-spacer signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226037pATB_146Plasmid encodes an AT-FP with OprF-spacer signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226038pATB_151Plasmid encodes an AT-FP with YeeJ signal peptide, epitope linker, YeeJ translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226039pATB_152Plasmid encodes an AT-FP with YeeJ signal peptide, flex linker, YeeJ translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226040pATB_153Plasmid encodes an AT-FP with YeeJ signal peptide, rigid linker, YeeJ translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226041pATB_211Plasmid encodes an AT-FP with CtxB signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226042pATB_212Plasmid encodes an AT-FP with CtxB signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226043pATB_213Plasmid encodes an AT-FP with CtxB signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226044pATB_214Plasmid encodes an AT-FP with CtxB signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226045pATB_215Plasmid encodes an AT-FP with CtxB signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226046pATB_216Plasmid encodes an AT-FP with CtxB signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226047pATB_221Plasmid encodes an AT-FP with OprF signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226048pATB_222Plasmid encodes an AT-FP with OprF signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226049pATB_223Plasmid encodes an AT-FP with OprF signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226050pATB_224Plasmid encodes an AT-FP with OprF signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226051pATB_225Plasmid encodes an AT-FP with OprF signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226052pATB_226Plasmid encodes an AT-FP with OprF signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226053pATB_231Plasmid encodes an AT-FP with CtxB-spacer signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226054pATB_232Plasmid encodes an AT-FP with CtxB-spacer signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226055pATB_233Plasmid encodes an AT-FP with CtxB-spacer signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226056pATB_234Plasmid encodes an AT-FP with CtxB-spacer signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226057pATB_235Plasmid encodes an AT-FP with CtxB-spacer signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226058pATB_236Plasmid encodes an AT-FP with CtxB-spacer signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226059pATB_241Plasmid encodes an AT-FP with OprF-spacer signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226060pATB_242Plasmid encodes an AT-FP with OprF-spacer signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226061pATB_243Plasmid encodes an AT-FP with OprF-spacer signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226062pATB_244Plasmid encodes an AT-FP with OprF-spacer signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226063pATB_245Plasmid encodes an AT-FP with OprF-spacer signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226064pATB_246Plasmid encodes an AT-FP with OprF-spacer signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226065pATB_251Plasmid encodes an AT-FP with YeeJ signal peptide, epitope linker, YeeJ translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226066pATB_252Plasmid encodes an AT-FP with YeeJ signal peptide, flex linker, YeeJ translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226067pATB_253Plasmid encodes an AT-FP with YeeJ signal peptide, rigid linker, YeeJ translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226068pATB_311Plasmid encodes an AT-FP with CtxB signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226069pATB_312Plasmid encodes an AT-FP with CtxB signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226070pATB_313Plasmid encodes an AT-FP with CtxB signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226071pATB_314Plasmid encodes an AT-FP with CtxB signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226072pATB_315Plasmid encodes an AT-FP with CtxB signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226073pATB_316Plasmid encodes an AT-FP with CtxB signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226074pATB_321Plasmid encodes an AT-FP with OprF signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226075pATB_322Plasmid encodes an AT-FP with OprF signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226076pATB_323Plasmid encodes an AT-FP with OprF signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226077pATB_324Plasmid encodes an AT-FP with OprF signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226078pATB_325Plasmid encodes an AT-FP with OprF signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226079pATB_326Plasmid encodes an AT-FP with OprF signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226080pATB_331Plasmid encodes an AT-FP with CtxB-spacer signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226081pATB_332Plasmid encodes an AT-FP with CtxB-spacer signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226082pATB_333Plasmid encodes an AT-FP with CtxB-spacer signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226083pATB_334Plasmid encodes an AT-FP with CtxB-spacer signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226084pATB_335Plasmid encodes an AT-FP with CtxB-spacer signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226085pATB_336Plasmid encodes an AT-FP with CtxB-spacer signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226086pATB_341Plasmid encodes an AT-FP with OprF-spacer signal peptide, epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226087pATB_342Plasmid encodes an AT-FP with OprF-spacer signal peptide, flex linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226088pATB_343Plasmid encodes an AT-FP with OprF-spacer signal peptide, rigid linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226089pATB_344Plasmid encodes an AT-FP with OprF-spacer signal peptide, ∆epitope linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226090pATB_345Plasmid encodes an AT-FP with OprF-spacer signal peptide, ∆epitope∆β1 linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226091pATB_346Plasmid encodes an AT-FP with OprF-spacer signal peptide, ∆epitope∆β1∆connecting region linker, EhaA translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226092pATB_351Plasmid encodes an AT-FP with YeeJ signal peptide, epitope linker, YeeJ translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226093pATB_352Plasmid encodes an AT-FP with YeeJ signal peptide, flex linker, YeeJ translocation unit enabling the display of a passenger of interest at the bacterial cell surface
226094pATB_353Plasmid encodes an AT-FP with YeeJ signal peptide, rigid linker, YeeJ translocation unit enabling the display of a passenger of interest at the bacterial cell surface
235136pATB_1yzThe expression of 27 autotransporter fusion protein variants is set under control of the ParaBAD promoter.
235137pATB_2yzThe expression of 27 autotransporter fusion protein variants is set under control of the PrhaBAD promoter.
235138pATB_3yzThe expression of 27 autotransporter fusion protein variants is set under control of the pATB-PRox306 promoter.

Antibodies from Article