T7 Primase-helicase

T7 primase-helicase is a multifunctional protein complex that unwinds DNA and initiates synthesis of new strands during bacteriophage T7 genome replication. Powered by ATP hydrolysis, its helicase activity moves along single-stranded DNA to separate the duplex, while the primase activity recognizes specific DNA sequences and produces short RNA primers for DNA polymerase extension. This coordinated mechanism links template unwinding with primer formation, supporting efficient leading- and lagging-strand synthesis. In genetics research, T7 primase-helicase provides a tractable model for studying replication-fork dynamics, protein cooperation, and enzyme specificity, and it supports biochemical assays of DNA replication and genome maintenance.

T7 Primase-helicase - Related Videos

Education

JoVE Core - Molecular Biology

DNA Helicases

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2020

DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...

Research

JoVE Journal - Genetics

Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins

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Cited by 3 •

2017

We describe sensitive, gel-based discontinuous assays to examine the kinetics of lagging-strand initiation using the replication proteins of bacteriophage T7.

DNA Sequence Recognition by DNA Primase Using High-Throughput Primase Profiling

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Cited by 3 •

2019

Protein binding microarray (PBM) experiments combined with biochemical assays link the binding and catalytic properties of DNA primase, an enzyme that synthesizes RNA primers on template DNA. This method, designated as high-throughput primase profiling (HTPP), can be used to reveal DNA-binding patterns of a variety of enzymes.

Helicase Activity Measurement of a Target Protein Using Biotin-Labeled RNA Duplexes

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2025

In this video, we demonstrate the procedure to determine the helicase activity of a target protein to unwind the biotin-labeled dsRNA substrate. The activity of the enzyme was identified by analyzing the electrophoretic mobility shift, followed by a chemiluminescence assay using chemiluminescent enzyme-conjugated streptavidin.

Generation of GFP-Tagged Recombinant Respiratory Syncytial Virus Using a T7 Expression System

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2026

Source: Bouillier, C. et. al., Amplification, and Titration of Recombinant Respiratory Syncytial Viruses. J. Vis. Exp. (2019)This video demonstrates the generation of a green fluorescent protein (GFP)-expressing recombinant respiratory syncytial virus using a T7-based reverse genetics system. The co-transfection of plasmids encoding viral proteins and a GFP-tagged genome leads to protein expression and initiates virus rescue.

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