Table 1 Genes and gene cassettes that were integrated according to pAIO protocol.

From: Fast and antibiotic free genome integration into Escherichia coli chromosome

Target protein

Description of target protein

Cassette length (bp)

Integration strain

Number of positive integrants out of 16 screened colonies

GFPmut3.1

Model protein

1367

B::I-SceIRS + pAIO

16

GFPmut3.1-KanR

Model protein with kanamycin resistance protein

2748

B::I-SceIRS + pAIO

14

ß-Lactamase

Ampicillin resistance protein

1392

B::I-SceIRS + pAIO

16

6His-mCherry-dsbASSsfGFP

Combination of cytoplasmic (mCherry) and periplasmic model protein (sfGFP)

1890

B::I-SceIRS + pAIO

10

dsbASSsfGFP

Periplasmic model protein

1100

B::I-SceIRS + pAIO

8

6His-mCherry

Cytoplasmic model protein

1087

B::I-SceIRS + pAIO

10

ompASSTNFα

Tumor necrose factor

2489

B::I-SceIRS + pAIO

14

6His-BIWA4 (scFv)

Human single chain antibody fragment targeting cell-surface glycoprotein

900

B::I-SceIRS + pAIO

6

6His-hFGF-2

Fibroblast growth factor

2801

B::I-SceIRS + pAIO

5

ompASS6His-mTNFα

Tumor necrose factor with His-tag

2507

B::I-SceIRS + pAIO

12

Fab: dsbASS-BIBH1

Human antibody fragment targeting fibroblast

Activation protein with N-terminal dsbA-leader

2208

B::I-SceIRS + pAIO H::I-SceIRS + pAIO

4

2

Fab: ompASSBIBH1

Human antibody fragment targeting fibroblast

Activation protein with N-terminal ompA-leader

2208

B::I-SceIRS + pAIO H::I-SceIRS + pAIO

6

4

Fab: dsbASSBIWA4

Human antibody fragment targeting cell-surface glycoprotein with

N-terminal dsbA-leader

2157

B::I-SceIRS + pAIO H::I-SceIRS + pAIO

9

1

Fab: ompASSBIWA4

Human antibody fragment targeting cell-surface glycoprotein with

N-terminal ompA-leader

2157

B::I-SceIRS + pAIO H::I-SceIRS + pAIO

10

2

Fab: dsbASSFTN2

Human antibody fragment targeting tumor necrose factor with N-terminal dsbA-leader

2172

B::I-SceIRS + pAIO H::I-SceIRS + pAIO

2

3

Fab: ompASSFTN2

Human antibody fragment targeting tumor necrose factor with N-terminal ompA-leader

2172

B::I-SceIRS + pAIO H::I-SceIRS + pAIO

11

2

  1. sfGFP…superfolder GFP, 6His…N-terminal His-tag, dsbASS…signal peptide of periplasmic thiol:disulfide interchange protein DsbA, ompASS…signal peptide of periplasmic outer membrane protein A, B… BL21(DE3), H…HMS174(DE3).
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