Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Coordination of Gene Expression Processes in Bacteria01:29

Coordination of Gene Expression Processes in Bacteria

The DNA replication, transcription, and translation processes are intricately coupled in bacteria, allowing efficient gene expression and rapid protein synthesis. While this physical and functional coordination is advantageous, it introduces challenges that bacteria overcome through specific regulatory mechanisms.Coupling of Replication, Transcription, and TranslationThe coupling of replication, transcription, and translation is a hallmark of bacterial gene expression. As the replisome unwinds...
Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Force-moment mechanics of wiggling in connector insertion.

Nature communications·2026
Same author

Limits and mechanisms of honey bee colonial thermoregulation in the heat.

The Journal of experimental biology·2026
Same author

Amplicon and metagenomic data from fumarole-associated geothermal features of Hawai'i.

Scientific data·2026
Same author

Hospital-Based Cross-Sectional Study of <i>Burkholderia pseudomallei</i> Seroreactivity Among Febrile Patients in Northernmost Vietnam: Near-Neighbor Bloodstream Isolates, Environmental Correlates, and Spatial Clustering.

GeoHealth·2026
Same author

Spatiotemporal dynamics and environmental trends of reported human leptospirosis in Sri Lanka, 2007-2024.

Frontiers in public health·2026
Same author

<i>N</i>-acyl-homoserine lactone-based quorum sensing beyond canonical lineages: insights from Actinomycetota.

Frontiers in microbiology·2026
Same journal

Complete sequencing of medaka genomes reveals the architecture of centromeric satellites, giant mobile elements, and sex chromosomes.

Genome research·2026
Same journal

Convergence and conflict among telomere specialized transposons across 60 million years of Drosophilid evolution.

Genome research·2026
Same journal

A unified analysis of cell type- and trajectory-associated pathways in single-cell data using Phoenix.

Genome research·2026
Same journal

Resf1 is required for proper placental development and configuration of trophoblast cell-specific heterochromatin.

Genome research·2026
Same journal

Telomere-driven replicative crisis is driven by large-scale changes in genomic architecture.

Genome research·2026
Same journal

Spatially informed reference-free cell-type deconvolution for spatial transcriptomics with SpatialCD.

Genome research·2026
See all related articles

Related Experiment Video

Updated: Jun 2, 2026

Transcriptome Analysis of Single Cells
07:27

Transcriptome Analysis of Single Cells

Published on: April 25, 2011

Transcript amplification from single bacterium for transcriptome analysis.

Yun Kang1, Michael H Norris, Jan Zarzycki-Siek

  • 1Department of Microbiology, University of Hawaii at Manoa, Honolulu, Hawaii 96822, USA.

Genome Research
|May 4, 2011
PubMed
Summary
This summary is machine-generated.

This study introduces a new total transcript amplification method for single prokaryotic cells. This technique enables robust transcriptome analysis from individual bacteria, overcoming previous limitations.

More Related Videos

A Fast and Reliable Pipeline for Bacterial Transcriptome Analysis Case study: Serine-dependent Gene Regulation in Streptococcus pneumoniae
10:18

A Fast and Reliable Pipeline for Bacterial Transcriptome Analysis Case study: Serine-dependent Gene Regulation in Streptococcus pneumoniae

Published on: April 25, 2015

Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics
13:51

Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics

Published on: February 18, 2009

Related Experiment Videos

Last Updated: Jun 2, 2026

Transcriptome Analysis of Single Cells
07:27

Transcriptome Analysis of Single Cells

Published on: April 25, 2011

A Fast and Reliable Pipeline for Bacterial Transcriptome Analysis Case study: Serine-dependent Gene Regulation in Streptococcus pneumoniae
10:18

A Fast and Reliable Pipeline for Bacterial Transcriptome Analysis Case study: Serine-dependent Gene Regulation in Streptococcus pneumoniae

Published on: April 25, 2015

Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics
13:51

Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics

Published on: February 18, 2009

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genomics

Background:

  • Total transcript amplification (TTA) is established for eukaryotic cells but challenging for prokaryotes due to limited starting material and lack of poly(A)-tails.
  • Prokaryotic mRNA can be polycistronic, adding complexity to transcriptome analysis.
  • Single-cell analysis provides high-resolution insights into cellular heterogeneity and responses.

Purpose of the Study:

  • To develop and validate a novel method for total transcript amplification (TTA) from single prokaryotic cells.
  • To assess the reliability and accuracy of the TTA method for transcriptome analysis in bacteria.
  • To investigate the transcriptional response of *Burkholderia thailandensis* to glyphosate exposure at the single-cell level.

Main Methods:

  • Developed a novel single-bacterium total transcript amplification (TTA) method.
  • Utilized *Burkholderia thailandensis* as a model organism exposed to subinhibitory glyphosate concentrations.
  • Assessed TTA method performance using microarrays, comparing results to larger-scale samples.
  • Validated single-cell data using *lacZ* and *gfp* reporter-gene fusions.
  • Confirmed sequence integrity and absence of contamination via Sanger sequencing.

Main Results:

  • The TTA method demonstrated low fold-change bias (less than twofold) and high correlation (R ≈ 0.87-0.89) compared to nonamplified samples.
  • Gene drop-out rates were low (4%-6% of detectable genes).
  • Microarray analysis suggested *B. thailandensis* modulates gene expression to manage amino acid pools when exposed to glyphosate.
  • Reporter gene assays confirmed the validity of single-cell microarray data.
  • Sanger sequencing confirmed the absence of contamination in TTA products.

Conclusions:

  • The novel TTA method is effective for analyzing the transcriptome of single prokaryotic cells.
  • This method overcomes challenges associated with prokaryotic RNA and limited starting material.
  • The findings suggest potential mechanisms for *B. thailandensis* adaptation to glyphosate stress.
  • The developed method opens possibilities for single-cell RNA sequencing in bacteria.