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Related Concept Videos

DNA-only Transposons02:57

DNA-only Transposons

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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...
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Urinary Tract Calculi I: Introduction01:28

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Renal calculi, or kidney stones, are solid deposits of minerals and salts formed inside the kidneys. In medical terminology, "calculus" refers to the stone itself, while "lithiasis" describes the process of stone formation. Depending on their location within the urinary system, these stones may be classified as either urolithiasis, when situated within the urinary tract, or nephrolithiasis, when located within the kidneys. Each term signifies the specific impact of the stone.Predisposition...
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Urinary Tract Infection I: Introduction01:26

Urinary Tract Infection I: Introduction

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Urinary tract infections (UTIs) impact various parts of the urinary system, including the kidneys, ureters, bladder, and urethra. These infections are generally bacterial, with Escherichia coli being the most common causative agent, often originating from the gastrointestinal tract. However, other bacteria, such as Staphylococcus saprophyticus, Klebsiella pneumoniae, and Proteus mirabilis, are also known to cause UTIs. The type, location, and underlying complexity of the UTI guide both...
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Urinary Tract Infection II: Pathophysiology01:25

Urinary Tract Infection II: Pathophysiology

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The pathophysiology of urinary tract infections (UTIs) encompasses several progressive stages, beginning with bacterial colonization and culminating in potential systemic complications if untreated. UTIs are primarily initiated by bacteria, such as Escherichia coli, which often originate from the gastrointestinal tract and migrate to the urinary system through the periurethral area. This migration can occur via several routes, including improper hygiene practices, sexual activity, or...
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Urinary Tract Calculi V: Nursing Management01:28

Urinary Tract Calculi V: Nursing Management

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AssessmentSubjective Data: Obtain a detailed health history, including any recent or chronic urinary tract infections, periods of immobilization, previous episodes of renal calculi, and medical conditions such as gout, benign prostatic hyperplasia, or hyperparathyroidism. Review the medication history for drugs that may influence stone formation, including allopurinol, analgesics, loop diuretics, or thiazide diuretics. Document the use of long-term indwelling catheters and any past surgical...
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Urinary Tract Calculi VI: Surgical Management01:25

Urinary Tract Calculi VI: Surgical Management

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Procedures for Kidney StonesMedical intervention is necessary when kidney stones or renal calculi are too large to pass spontaneously (typically greater than 5 millimeters) when stones are accompanied by symptomatic infection (such as fever or pyelonephritis), when they impair kidney function, or when they cause persistent symptoms like severe pain, nausea, or urinary retention. Additionally, patients with only one kidney or those who cannot be treated with medical management also require...
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Related Experiment Video

Updated: Jan 22, 2026

Ultrasonography of the Adult Male Urinary Tract for Urinary Functional Testing
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Transposon Insertion Site Sequencing in a Urinary Tract Model.

Valerie S Forsyth1, Harry L T Mobley1, Chelsie E Armbruster2

  • 1Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 17, 2019
PubMed
Summary

Transposon sequencing (Tn-seq) enables quantitative analysis of mutant abundance during infection. This method efficiently determines gene fitness contributions in Proteus mirabilis.

Keywords:
Illumina sequencingSouthern blotTn-seqTransposon sequencingUrinary tract infection

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Area of Science:

  • Microbiology
  • Genomics
  • Infectious Diseases

Background:

  • Transposon sequencing (Tn-seq) is a powerful technique for large-scale genetic analysis.
  • It combines mutant library generation with next-generation sequencing to assess gene function.
  • Traditional methods for analyzing mutant pools are less efficient and comprehensive.

Purpose of the Study:

  • To describe a method for generating a genome-saturating transposon mutant library in Proteus mirabilis.
  • To outline the process for preparing Tn-seq libraries for Illumina sequencing.
  • To detail the downstream analysis for estimating gene fitness during infection.

Main Methods:

  • Generation of a genome-saturating transposon mutant library in Proteus mirabilis.
  • Determination of optimal mutant library size for experimental infection inoculation.
  • Preparation of transposon insertion junctions for high-throughput sequencing.
  • Bioinformatic analysis of sequencing reads to quantify mutant abundance and infer gene fitness.

Main Results:

  • A robust Tn-seq methodology was established for Proteus mirabilis.
  • The study determined optimal parameters for experimental infection models using Tn-seq.
  • Analysis provided quantitative data on the fitness contribution of genes during infection.

Conclusions:

  • Tn-seq is an effective approach for high-throughput fitness profiling in Proteus mirabilis.
  • This method significantly advances the study of microbial pathogenesis and gene essentiality.
  • The described protocol facilitates comprehensive genetic analysis in bacterial pathogens.