Dulbecco’s modified Eagle medium (DMEM),
fetal bovine serum (FBS),l -glutamine, penicillin/streptomycin
(P/S), Opti-MEM I (OMEM), Simple Blue Stain, Novex 4–12% Tris-glycine
SDS-PAGE gels, Novex Sharp Unstained Protein Standard, GeneRuler 1
kb Plus DNA Ladder, LB Medium Dehydrated Capsules, and the Phusion
High-Fidelity DNA Polymerase were all purchased from Thermo Fisher
Scientific. His-tagged Pfu X7 DNA Polymerase was prepared in-house
by immobilized metal affinity chromatography for routine PCR-based
bacterial colony screening. XL10-Gold Ultracompetent cells were acquired
from Agilent Technologies, Inc. Note XL10-Gold cells have a mutation
in the lacI gene promoter (lacIq), resulting in higher Lac repressor levels.57 (link) SIGMAFAST EDTA-free Protease Inhibitor cocktail
tablets and IPTG were obtained from Sigma-Aldrich. DpnI, TransIT-LT1 transfection reagent, and 2′-(4-methylumbelliferyl)-α-d -N-acetylneuraminic acid (MUNANA) were obtained
from New England Biolabs, Mirus Bio, and Cayman Chemicals, respectively.
Specific-pathogen-free (SPF) eggs and turkey red blood cells (TRBCs)
were purchased from Charles River Labs and the Poultry Diagnostic
and Research Center (Athens, GA), respectively. All cloning and screening
primers (Table S1 ) used in this study were
synthesized by Integrated DNA Technologies.
fetal bovine serum (FBS),
(P/S), Opti-MEM I (OMEM), Simple Blue Stain, Novex 4–12% Tris-glycine
SDS-PAGE gels, Novex Sharp Unstained Protein Standard, GeneRuler 1
kb Plus DNA Ladder, LB Medium Dehydrated Capsules, and the Phusion
High-Fidelity DNA Polymerase were all purchased from Thermo Fisher
Scientific. His-tagged Pfu X7 DNA Polymerase was prepared in-house
by immobilized metal affinity chromatography for routine PCR-based
bacterial colony screening. XL10-Gold Ultracompetent cells were acquired
from Agilent Technologies, Inc. Note XL10-Gold cells have a mutation
in the lacI gene promoter (lacIq), resulting in higher Lac repressor levels.57 (link) SIGMAFAST EDTA-free Protease Inhibitor cocktail
tablets and IPTG were obtained from Sigma-Aldrich. DpnI, TransIT-LT1 transfection reagent, and 2′-(4-methylumbelliferyl)-α-
from New England Biolabs, Mirus Bio, and Cayman Chemicals, respectively.
Specific-pathogen-free (SPF) eggs and turkey red blood cells (TRBCs)
were purchased from Charles River Labs and the Poultry Diagnostic
and Research Center (Athens, GA), respectively. All cloning and screening
primers (
synthesized by Integrated DNA Technologies.