Construction of vaccine from Lactococcus lactis bacteria using Aeromonas hydrophila virulent Aerolysin gene

Authors

  • Foo Hai Ling
  • Hassan Mohd Davoud
  • Raha Abd Rahim
  • Son Radu
Abstract:

 In this study the forward and reverse primers were designated to amplify the segments (~250 bps and ~650 bps) of the gene coding domains 1 and 4 of aerolysin of Aeromonas hydrophila. These two domains are involved in pathogenesis of the aerolysin gene. Sequences for two restriction enzymes, Pst I and Hind III, were included in the forward and reverse primers respectively. These restriction enzyme sites were used because they are not present within the genes of interest but are available in the multiple cloning sites of plasmid pNZ8048. Amplified PCR products were analyzed with 1% agarose gel electrophoresis and results showed that amplifications were very specific. In comparison with the DNA marker, the sizes of the amplified PCR products were determined to be approximately ~250 bps and ~650 bps respectively. PCR products were then purified by the DNA purification kit, digested with REs and ligated with linearised pNZ8048 plasmid using T4 DNA ligase. Transformation of Lactococcus lactis NZ9000 cells was performed by the electroporation method. Verification for cloning of virulent genes was performed by REs digestion and also DNA sequencing. Since several antigens (bacterial and viral) and cytokines have been efficiently produced in L. lactis, constructing and expression and utilization of recombinant L. lactis harboring the aerolysin domains (virulent) genes from A. hydrophila may induce production of antibodies in fish against this pathogen.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

Cloning of EprA1 gene of Aeromonas hydrophila in Lactococcus lactis

Bacterial-based systems as live vectors for the delivery of heterologous antigens offer a number of advantages as vaccination strategies. Developments in genetic engineering have given Gram-positive lacticacid bacteria (LAB) the advantage of being used as a host expression system for antigen delivery to inducethe immune response. A fragment containing the full length of the “eprA1” ...

full text

The cytotoxic enterotoxin of Aeromonas hydrophila is aerolysin.

The channel-forming toxin aerolysin was identified 25 years ago by Bernheimer and Avigad (1), who gave the protein its name. Aerolysin was purified by Buckley et al. (2), and its structural gene, named aerA, was cloned and sequenced almost simultaneously by two groups (6, 7). Since then more than 50 articles describing the expression, secretion, and properties of aerolysin have appeared, and ae...

full text

Determination of the optimal conditions of cloning Aerolysin gene from the common carp pathogen Aeromonas hydrophila in Escherichia coli BL21

Aeromonas hydrophila is a gram-negative bacterium which associated with gastrointestinal diseases and septicaemia. This pathogenic bacterium has several virulence factors ranging from pili to the excreted protein which called (Aerolysin) with minor and major effects, respectively. Additionally, Aeromonas hydrophila is a widely distributed bacterium that commonly causes ulcers in cyprinid fish s...

full text

Detection of the aerolysin gene in Aeromonas hydrophila by the polymerase chain reaction.

Synthetic oligonucleotide primers were used in a polymerase chain reaction (PCR) technique to detect the gene for aerolysin in strains of Aeromonas hydrophila and to screen for identical genes in A. caviae, A. sobria, and A. veronii isolated from patients with diarrheal disease. Primers targeted a 209-bp fragment of the aer gene coding for the beta-hemolysin and detected template DNA only in th...

full text

Cloning, expression and characterization of aerolysin from Aeromonas hydrophila in Escherichia coli.

Aerolysin is a toxin (protein in nature) secreted by the strains of Aeromonas spp. and plays an important role in the virulence of Aeromonas strains. It has also found several applications such as for detection of glycosylphosphatidylinositol (GPI)-anchored proteins etc. A. hydrophila is a ubiquitous Gram-negative bacterium which causes frequent harm to the aquaculture. To obtain a significant ...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 10  issue None

pages  143- 154

publication date 2011-01

By following a journal you will be notified via email when a new issue of this journal is published.

Keywords

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023