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Related Concept Videos

Mechanism of Conjugation01:19

Mechanism of Conjugation

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...
Conjugation01:19

Conjugation

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...
DNA Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
Gene Conversion02:08

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
Phase II Conjugation Reactions: Overview01:14

Phase II Conjugation Reactions: Overview

Conjugation, a key component of phase II biotransformation reactions, is a vital process in drug detoxification. It involves transferring endogenous substances like glucuronic acid, sulfate, and glycine to drugs or their metabolites formed in phase I reactions. These conjugation reactions, often catalyzed by specific enzymes, transform potentially harmful metabolites into inactive, water-soluble forms easily excreted in urine or bile. By enhancing polarity and eliminating pharmacological...
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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Related Experiment Video

Updated: Jul 1, 2026

Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation
10:41

Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation

Published on: January 4, 2017

DNA conjugation by Staudinger ligation.

Samuel H Weisbrod1, Anna Baccaro, Andreas Marx

  • 1Department of Chemistry, University of Konstanz, 78457, Konstanz, Germany. samuel.weisbrod@uni-konstanz.de

Nucleic Acids Symposium Series (2004)
|September 9, 2008
PubMed
Summary
This summary is machine-generated.

Researchers synthesized modified nucleotides with terminal azides for DNA applications. These nucleotides function in primer extension and polymerase chain reactions (PCR), enabling subsequent Staudinger ligation for attaching tags like biotin.

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Area of Science:

  • Nucleic acid chemistry
  • Bioconjugation chemistry
  • Molecular biology

Background:

  • Modified nucleotides are essential tools in molecular biology.
  • Developing efficient methods for nucleotide modification and subsequent bioconjugation is crucial for expanding their applications.

Purpose of the Study:

  • To synthesize modified 2'-deoxyuridine triphosphates and 2'-deoxy-7-deazaadenosine with terminal azides.
  • To evaluate the steric influence of linkers on nucleotide incorporation and Staudinger ligation.
  • To demonstrate the utility of these modified nucleotides in primer extension and PCR.

Main Methods:

  • Chemical synthesis of azide-modified nucleotides.
  • Incorporation studies using primer extension and polymerase chain reactions (PCR) with Pwo DNA polymerase.
  • Staudinger ligation for bioconjugation of azide-labeled DNA products with phosphines.

Main Results:

  • Successfully synthesized three modified nucleotides with terminal azides.
  • Demonstrated complete replacement of natural nucleotides in primer extension and PCR.
  • Achieved successful Staudinger ligation for conjugating biotin as an affinity tag to azide-labeled DNA.

Conclusions:

  • The synthesized modified nucleotides are compatible with enzymatic DNA synthesis.
  • Staudinger ligation provides a versatile platform for post-synthetic modification of DNA.
  • These modified nucleotides offer a valuable tool for various molecular biology applications, including affinity-based purification.