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Related Concept Videos

Bacterial Transformation01:33

Bacterial Transformation

58.4K
In 1928, bacteriologist Frederick Griffith worked on a vaccine for pneumonia, which is caused by Streptococcus pneumoniae bacteria. Griffith studied two pneumonia strains in mice: one pathogenic and one non-pathogenic. Only the pathogenic strain killed host mice.
Griffith made an unexpected discovery when he killed the pathogenic strain and mixed its remains with the live, non-pathogenic strain. Not only did the mixture kill host mice, but it also contained living pathogenic bacteria that...
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Transduction01:16

Transduction

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Among the three main modes of HGT—transformation, conjugation, and transduction—transduction is unique in that it is mediated by bacteriophages, or bacterial viruses.Transduction occurs in two ways. Generalized transduction occurs during the lytic cycle of a bacteriophage infection. In this process, bacteriophages infect bacterial cells, replicate within them, and ultimately cause cell lysis, releasing newly assembled virions. Occasionally, random fragments of the bacterial genome...
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Mechanism of Conjugation01:19

Mechanism of Conjugation

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Bacterial conjugation is a mechanism of horizontal gene transfer that enables the exchange of genetic material between bacterial cells through direct contact. This process is facilitated by a donor cell carrying a conjugative plasmid, which encodes genes necessary for pilus formation, DNA replication, and transfer. The conjugative plasmid plays a central role in initiating and executing the transfer of genetic material.The tra region of the conjugative plasmid encodes proteins responsible for...
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Transformation01:26

Transformation

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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...
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Conjugation01:19

Conjugation

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Conjugation is a form of horizontal gene transfer that primarily occurs in bacteria and some archaea, promoting genetic diversity and adaptation. Bacteria can acquire resistance genes through conjugative plasmids, allowing them to survive antibiotic treatments that would otherwise be lethal. This process involves direct contact between cells through specialized structures such as the sex pilus and is mediated by conjugative plasmids, including the F (fertility) factor.Conjugation requires...
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Conjugative DNA Transfer From E. coli to Transformation-Resistant Lactobacilli.

Sara Samperio1, Dolores L Guzmán-Herrador1, Rigoberto May-Cuz1

  • 1Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC), Universidad de Cantabria-CSIC-SODERCAN, Santander, Spain.

Frontiers in Microbiology
|March 1, 2021
PubMed
Summary
This summary is machine-generated.

Bacterial conjugation is a new genetic modification tool for lactic acid bacteria (LAB), including difficult-to-transform species. This method successfully introduced genetic material into various LAB and even Staphylococcus epidermidis, expanding genetic engineering possibilities.

Keywords:
LactobacillusStaphylococcus epidermidisbacterial conjugationlactic acid bacteriaplasmid R388plasmid RP4

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Creation of a Dense Transposon Insertion Library Using Bacterial Conjugation in Enterobacterial Strains Such As Escherichia Coli or Shigella flexneri
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Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation
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Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Lactic acid bacteria (LAB), including the genus formerly known as Lactobacillus, are crucial in the food industry and as probiotics.
  • Genetic modification tools for LAB are limited, especially for wild-type and non-model strains.
  • Developing new genetic tools is essential for harnessing the full potential of LAB in food and therapeutic applications.

Purpose of the Study:

  • To evaluate bacterial conjugation as a novel method for genetic modification of diverse LAB species.
  • To assess the efficiency of conjugation systems (RP4 and R388) in transferring plasmids into various lactobacilli and other Gram-positive bacteria.
  • To establish a reliable protocol for genetic manipulation of strains previously resistant to transformation.

Main Methods:

  • Utilized bacterial conjugation with mobilizable shuttle plasmids from donor Escherichia coli strains.
  • Employed RP4 and R388 conjugative systems for DNA transfer.
  • Tested conjugation efficiency across multiple Lactocaseibacillus casei strains, Lentilactobacillus parabuchneri, and Staphylococcus epidermidis.
  • Confirmed transconjugants via oriT presence and 16S rRNA gene sequencing.

Main Results:

  • Successfully achieved bacterial conjugation in all tested Lactocaseibacillus casei strains, including natural isolates.
  • Demonstrated successful conjugation into Lentilactobacillus parabuchneri, a species with no previously reported transformation protocol.
  • Transconjugants were also obtained in Staphylococcus epidermidis, a common laboratory contaminant.
  • Confirmed the efficacy of both RP4 and R388 conjugative systems for plasmid mobilization into S. epidermidis.

Conclusions:

  • Bacterial conjugation is a versatile and effective tool for genetic modification of a wide range of lactic acid bacteria.
  • This method overcomes limitations of existing transformation protocols, enabling genetic engineering of previously intractable strains.
  • The established conjugation protocol holds promise for genetic manipulation of other Gram-positive microorganisms resistant to transformation, broadening biotechnological applications.