Related Experiment Video

Updated: Sep 28, 2026

Methods for Electroporation and Transformation Confirmation in Limosilactobacillus reuteri DSM20016
11:04

Methods for Electroporation and Transformation Confirmation in Limosilactobacillus reuteri DSM20016

Published on: June 23, 2023

Survey of restriction-modification systems and transformation in Mannheimia haemolytica and Pasteurella trehalosi

A E Hill1, F A Lainson

  • 1Moredun Research Institute, International Research Centre, Pentlands Science Park, Bush Loan, Penicuik, Midlothian, Edinburgh, Scotland EH26 0PZ, UK. a.hill@hw.ac.uk

Veterinary Microbiology
|December 19, 2002
PubMed

Insights

Genetic transformation of Mannheimia haemolytica strains was achieved using electroporation. This overcomes a major hurdle in molecular studies, even with existing restriction modification systems present in the bacteria.

Area of Science:

  • Bacteriology
  • Molecular Biology
  • Genetics

Background:

  • Mannheimia (Pasteurella) haemolytica exhibits resistance to genetic transformation, hindering molecular research.
  • This resistance is often attributed to the presence of restriction modification systems within the bacterial strains.

Purpose of the Study:

  • To overcome the genetic transformation barrier in Mannheimia haemolytica.
  • To establish a reliable method for transforming various serotypes of M. haemolytica and Pasteurella trehalosi.

Main Methods:

  • Electroporation was employed as the transformation technique.
  • A recombinant vector derived from the M. haemolytica A1 serotype plasmid pNSF2176 was utilized.
  • Polymerase Chain Reaction (PCR) was used to detect restriction enzyme genes.

Main Results:

  • Successful genetic transformation was achieved in representative strains of 12 M. haemolytica serotypes and four Pasteurella trehalosi serotypes.
  • Transformation occurred despite the identified presence of type I (phaI) and type II (hsdM) restriction enzyme genes in the M. haemolytica strains.

Conclusions:

  • Electroporation provides an effective method for genetic transformation of diverse M. haemolytica and P. trehalosi serotypes.
  • The presence of restriction modification systems does not preclude successful transformation using this method, opening avenues for further molecular studies.

More Related Videos

Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus10:39

Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus

Published on: March 10, 2017

Purification of a High Molecular Mass Protein in Streptococcus mutans09:51

Purification of a High Molecular Mass Protein in Streptococcus mutans

Published on: September 14, 2019

Related Concept Videos

Transformation01:26

Transformation

Microbial communities are dynamic environments where cell lysis releases free DNA into the surroundings. Other cells can take up this extracellular DNA through a process known as transformation.When a cell incorporates this foreign DNA into its genome, resulting in genetic modification, the process is known as transformation. Cells capable of this process are termed competent. Competence can be natural, as observed in certain bacteria and archaea, or artificially induced in the...
Clinical Significance of Antibiotic Resistance01:25

Clinical Significance of Antibiotic Resistance

Methicillin-resistant Staphylococcus aureus (MRSA) presents a critical public health threat, arising from its capacity to resist β-lactam antibiotics due to acquisition of the mecA gene within the staphylococcal cassette chromosome mec (SCCmec). This gene encodes penicillin-binding protein 2a (PBP2a), which impairs binding efficacy of methicillin and other β-lactams. MRSA has evolved into distinct clonal lineages impacting humans and animals alike, reinforcing its significance within the One...
Regulation of Bacterial Virulence01:28

Regulation of Bacterial Virulence

Pathogenic bacteria employ a range of regulatory mechanisms to modulate the expression of virulence genes in response to environmental and host-derived signals. These mechanisms ensure that virulence factors are expressed only under favorable conditions, thereby optimizing infection and survival strategies.Mechanisms of Virulence RegulationKey regulatory strategies include:Two-Component Systems: These consist of a membrane-bound sensor kinase and a cytoplasmic response regulator. Environmental...