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Related Concept Videos

Transposons01:24

Transposons

Transposons, or "jumping genes," are small mobile genetic elements (MGEs) that range from 700 to 40,000 base pairs in length. They are found in all organisms and can move within the same chromosome or transfer to different chromosomes. In some cases, transposons can also jump between different host DNA molecules, such as plasmids or viruses, contributing to genetic variability.Barbara McClintock first discovered these mobile genetic elements in the 1940s while studying maize genetics, and she...
DNA-only Transposons02:57

DNA-only Transposons

DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
DNA Bacteriophages01:26

DNA Bacteriophages

Bacteriophages, or phages, are viruses that specifically infect bacteria, utilizing their genetic material to hijack host cellular machinery for replication. DNA bacteriophages employ single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA) genomes. These phages exhibit diverse replication strategies and host interactions, influencing their ecological roles and applications in biotechnology and medicine.ssDNA BacteriophagesssDNA phages, with their small genomes, utilize unique strategies to...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
Transduction01:16

Transduction

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 are...

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Updated: Jun 12, 2026

Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis
13:31

Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis

Published on: October 31, 2014

Phage transposon mutagenesis.

M Sloan Siegrist1, Eric J Rubin

  • 1Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 19, 2010
PubMed
Summary
This summary is machine-generated.

This study details PhiMycoMarT7 phage preparation and transduction for creating mycobacterial transposon mutant libraries. These libraries enable efficient forward and reverse genetic studies in mycobacteria.

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Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing
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Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing

Published on: July 7, 2020

Area of Science:

  • Microbiology
  • Genetics
  • Molecular Biology

Background:

  • Phage transduction offers a promising avenue for genetic manipulation in mycobacteria.
  • PhiMycoMarT7 is a specialized phage well-suited for transposon mutagenesis.

Purpose of the Study:

  • To provide a detailed protocol for preparing PhiMycoMarT7 phage.
  • To outline the procedure for mycobacterial transduction using PhiMycoMarT7.
  • To facilitate the generation of transposon mutant libraries for genetic studies.

Main Methods:

  • Preparation of PhiMycoMarT7 phage stocks, including reconstitution and purification.
  • Titering of phage stock to determine infectivity.
  • Transduction of mycobacteria with PhiMycoMarT7.
  • Isolation of individual transposon mutants.

Main Results:

  • A comprehensive protocol for PhiMycoMarT7 phage preparation and transduction is established.
  • Transposon mutant libraries in mycobacteria can be efficiently generated.
  • The protocol supports both forward and reverse genetic analyses.

Conclusions:

  • PhiMycoMarT7 phage transduction is a robust method for generating mycobacterial mutant libraries.
  • This protocol enables advanced genetic studies in mycobacteria, aiding in understanding gene function.