Renaturation and purification of ApxII toxin of Actinobacillus pleuropneumoniae.

Protein Expr Purif

Division of Bacterial Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.

Published: April 2007

ApxII toxin is the only Apx toxin that is produced by Actinobacillus pleuropneumoniae serotype 7. In order to determine whether the recombinant ApxII that derived from Escherichia coli (E. coli) expression is faithful to the natural ApxII so that can be used as additional component in vaccine preparation, the structure gene apxIIA of ApxII toxin was expressed in E. coli with prokaryotic expression vector pGEX-6p-1 (formed pGEX-6p-A). pGZRS-C which is A. pleuropneumoniae-E. coli shuttle vector pGZRS-38 expressing the post-transcriptional activation gene apxII C was co-expressed with pGEX-6p-A. The expression product of rApxII A formed inclusion. The inclusion protein was oxidized, refolded and restored hemolytic activity after denaturation, renaturation and purification. The result indicated that E. coli expressed recombinant ApxII toxin has good fidelity, which makes it possible to produce this valuable antigen for vaccine preparation or diagnosis.

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.pep.2006.10.026DOI Listing

Publication Analysis

Top Keywords

apxii toxin
16
renaturation purification
8
actinobacillus pleuropneumoniae
8
recombinant apxii
8
vaccine preparation
8
apxii
7
toxin
5
coli
5
purification apxii
4
toxin actinobacillus
4

Similar Publications

The ApxIVA protein belongs to a distinct class of a "clip and link" activity of Repeat-in-ToXin (RTX) exoproteins. Along with the three other pore-forming RTX toxins (ApxI, ApxII and ApxIII), ApxIVA serves as a major virulence factor of Actinobacillus pleuropneumoniae, the causative agent of porcine pneumonia. The gene encoding ApxIVA is located on a bicistronic operon downstream of the orf1 gene and is expressed exclusively under in vivo conditions.

View Article and Find Full Text PDF

Emerging resistance to florfenicol in Actinobacillus pleuropneumoniae isolates on two Italian pig farms.

Vet Microbiol

September 2024

Istituto Zooprofilattico Sperimentale Dell'umbria E Delle Marche 'Togo Rosati', Italy; Istituto Zooprofilattico Sperimentale Della Lombardia E Dell'emilia Romagna, Italy.

Article Synopsis
  • Actinobacillus pleuropneumoniae causes porcine pleuropneumonia, a serious lung infection in pigs, and is typically managed using antibiotics like phenicols.
  • This study identifies three florfenicol-resistant isolates of A. pleuropneumoniae from Italian pig farms, all harboring the floR gene and showing varying susceptibility levels.
  • Whole genome sequencing revealed moderate virulence traits and the presence of a novel plasmid, pAp-floR, which is closely related to another plasmid and shows stability without selective pressure, emphasizing the need to monitor resistance in this bacterium.
View Article and Find Full Text PDF

The ApxII toxin and the outer membrane lipoprotein (Oml) of Actinobacillus pleuropneumoniae are important vaccine antigens against porcine contagious pleuropneumonia (PCP), a prevalent infectious disease affecting the swine industry worldwide. Previous studies have reported the recombinant expression of ApxII and Oml in Escherichia coli; however, their yields were not satisfactory. Here, we aimed to enhance the production of ApxII and Oml by constructing a bicistronic expression system based on the widely used T7 promoter.

View Article and Find Full Text PDF

Genetic Diversity of Serovars in Hungary.

Vet Sci

September 2022

Department of Microbiology and Infectious Diseases, University of Veterinary Medicine, Hungária krt. 23-25, H-1143 Budapest, Hungary.

A total of 114 isolates from porcine hemorrhagic necrotic pleuropneumonia were characterized by the examination of biotype, serovar, antibiotic resistance genes, and genes of toxin production. Pulsed-field gel electrophoresis was used to analyze their genetic relationship, which identified 16 clusters. Serovar 2 (50 isolates), serovar 13 (25 isolates), serovar 9 (11 isolates), and serovar 16 (7 isolates) were the most frequent serovars.

View Article and Find Full Text PDF

N-terminus of flagellin enhances vaccine efficacy against Actinobacillus pleuropneumoniae.

BMC Vet Res

July 2022

Graduate Institute of Animal Vaccine Technology, College of Veterinary Medicine, National Pingtung University of Science and Technology, 1, Shuefu Road, Neipu, Pingtung, 91201, Taiwan.

Background: Flagellin elicits potent immune response and may serve as a vaccine adjuvant. We previously reported that the N-terminus of flagellin (residues 1-99, nFliC) is sufficient for vaccine efficacy enhancement against Pasteurella multocida challenge in chickens. In this study, we futher tested the adjuvancy of nFliC in a subunit vaccine against the pig pathogen Actinobacillus pleuropneumoniae in a mice model.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!