Recombinant BL21 with LngA Variants from ETEC E9034A Promotes Adherence to HT-29 Cells

Overview

Journal Pathogens

Date 2023 Feb 25

PMID 36839609

Authors

Karina Espinosa-Mazariego

Zeus Saldana-Ahuactzi

Sara A Ochoa

Bertha Gonzalez-Pedrajo

Miguel A Cevallos

Ricardo Rodriguez-Martinez

Mariana Romo-Castillo

Rigoberto Hernandez-Castro

Ariadnna Cruz-Cordova

Juan Xicohtencatl-Cortes

Affiliations

Soon will be listed here.

Abstract

Aim: This study aimed to determine whether . BL21 (ECBL) transformed with the operon lacking the gene (pE9034AΔ) and complemented with variants of ETEC clinical strains, as well as point substitutions, exhibited modified adherence to HT-29 cells.

Methods: A kanamycin cassette was used to replace the gene in the operon of the E9034A strain, and the construct was transformed into the ECBL strain. The pJET1.2 vector carrying genes with allelic variants was transformed into ECBLpE9034AΔ (ECBLΔ). The point substitutions were performed in the pJET vector.

Results: Bioinformatic alignment analysis of the LngA proteins showed hypervariable regions and clustered the clinical ETEC strains into three groups. Variations in amino acid residues affect the adherence percentages of recombinant ECBL strains with variants and site-specific mutations with HT-29 cells.

Conclusion: In this study, ECBL carrying the operon harboring variants of six clinical ETEC strains, as well as point substitutions, exerted an effect on the adherence of ECBL to HT-29 cells, thereby confirming the importance of the CS21 pilus in adherence.

Citing Articles

Genome Sequencing and Assembly of Enterotoxigenic E9034A: Role of LngA, CstH, and FliC in Intestinal Cell Colonization and the Release of the Proinflammatory Cytokine IL-8.

Rodriguez-Martinez R, Ochoa S, Valle-Rios R, Jaimes-Ortega G, Hernandez-Castro R, Mancilla-Rojano J Microorganisms. 2025; 13(2).

PMID: 40005742 PMC: 11858209. DOI: 10.3390/microorganisms13020374.

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