Table 1 E. coli strains, plasmids and primers used in this study

E. coli DH5α

RecA

Promega

E. coli MG1655

Wild type

Laboratory stock

E. coli MS04

MG1655 ΔpflB, ΔadhE, ΔfrdA, ΔxylFGH, gatC S184L, ΔmidarpA, Δreg 27.3 kb, ΔldhA, PpflB::pdc _Zm -adhB _Zm .

[5],[28]

E. coli BL21 (DE3)

F⁻ ompT hsdS _B (r _B ⁻ m _B ⁻) gal dcm (DE3)

Novagen

Plasmid

Description

Source

pTrc99A2

pTrc99A derivative modified for NdeI cloning instead of NcoI. Designed for IPTG-inducible expression of proteins under the hybrid trp/lac promoter. Amp^r.

[13]

pAIDABglC

pTrc99A derivative designed for cell surface display of BglC using the autotransporter AIDA-I. Amp^r.

[13]

pET-22b(+)

Designed for IPTG-inducible expression of proteins under the T7 promoter. Amp^r.

Novagen

pETflu

pET-22b(+) derivative that has cloned the flu gene.

This study

pAg43pol

pTrc99A derivative designed for cloning heterologous sequences for protein secretion using the autotransporter Ag43. Amp^r.

This study

pAg43BglC

pAg43pol derivative designed for cell surface display of BglC using the autotransporter Ag43. Amp^r.

This study

pNS6

pET-26b(+) derivative employed for intracellular heterologous production of BglC in E. coli.

[21]

^a Primer

Sequence 5′→3′

Source

DfluNde

CTAAGGAAAAGCATATGAAACGACATCTG

This study

RfluHind

CAGAGAGGCGAAGCTTCTGTCAGAAGGTC

This study

Pet38F (26)

CGATCTCGATCCCGCGAAATTAATAC

This study

RfluBNE

CGGTGAATTCCGCCATGGCAGATCTGTCAGCAGCCAGCACCGGGAG

This study

DfluKXB

GACCGGTACCCTCGAGCTGAACGGATCCATTGACCCCACG

This study