Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

The DNA Replication Fork

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork. Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...

The DNA Replication Fork

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork. Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...

Replication in Prokaryotes

Replication in Prokaryotes

DNA replication has three main steps: initiation, elongation, and termination. Replication in prokaryotes begins when initiator proteins bind to the single origin of replication (ori) on the cell's circular chromosome. Replication then proceeds around the entire circle of the chromosome in each direction from the two replication forks, resulting in two DNA molecules.
Many Proteins Work Together to Replicate the Chromosome
Replication is coordinated and carried out by a host of specialized...

Replication in Prokaryotes

Replication in Prokaryotes

Replication in Prokaryotes

Replication in Prokaryotes

Replication in Eukaryotes

Replication in Eukaryotes

In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
Many Proteins Orchestrate Replication at the Origin
Eukaryotic replication follows many of the same...

You might also read

Related Articles

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

Sort by

Same author

Cardiotropic AAV gene therapy for heart failure: a phase 1 trial.

Nature medicine·2025

Same author

IL-1ra gene therapy in equine osteoarthritis improves physiological, anatomical, and biological outcomes of joint degeneration.

Journal of the American Veterinary Medical Association·2024

Same author

Rationally engineered novel AAV capsids for intra-articular gene delivery.

Molecular therapy. Methods & clinical development·2024

Same author

Evaluation of an AAV9-mini-dystrophin gene therapy candidate in a rat model of Duchenne muscular dystrophy.

Molecular therapy. Methods & clinical development·2023

Same author

Improved understanding of biorisk for research involving microbial modification using annotated sequences of concern.

Frontiers in bioengineering and biotechnology·2023

Same author

Nicholas Muzyczka, PhD [1947-2023].

Human gene therapy·2023

Search research articles

Related Experiment Video

Updated: Jun 16, 2026

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Published on: August 21, 2016

Creating a novel origin of replication through modulating DNA-protein interfaces.

F Curtis Hewitt¹, R Jude Samulski

¹Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.

|January 29, 2010

Summary

Researchers identified key DNA-protein interactions governing adeno-associated virus (AAV) replication specificity between serotypes. This discovery advances understanding of viral DNA replication and AAV gene therapy applications.

More Related Videos

Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

Published on: September 11, 2017

Related Experiment Videos

Last Updated: Jun 16, 2026

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Published on: August 21, 2016

Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

Published on: September 11, 2017

Area of Science:

Virology
Molecular Biology
Genetics

Background:

DNA-protein interactions at replication origins are crucial but poorly understood in many higher eukaryotes and viruses.
Adeno-associated virus (AAV) uses a specific Replication protein (Rep) and origin for replication, yet serotype-specific binding mechanisms remain unclear.

Purpose of the Study:

To elucidate the molecular basis of DNA-protein specificity between different adeno-associated virus (AAV) serotypes and their replication origins.
To understand how the Rep protein recognizes and interacts with specific viral origins of replication.

Main Methods:

Generated chimeric and mutant origins between AAV2 and AAV5.
Performed in vivo replication assays.
Utilized structural modeling to analyze protein-DNA interactions.

Main Results:

Identified two independent DNA-protein interfaces critical for replicative specificity.
Determined three specific residues in AAV2 Rep are essential for cognate origin cleavage.
Characterized a unique interaction in AAV5 involving an extended binding element and a 49 amino acid Rep region with two DNA binding interfaces.

Conclusions:

Elucidated structure-function relationships at the AAV origin, enabling the creation of a novel recombinant origin and Rep protein.
The findings may enhance the safety and efficacy of AAV-based gene delivery vectors.
This research provides insights into conserved viral replication mechanisms and site-directed integration.