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Related Concept Videos

Single-Strand DNA Binding Proteins01:03

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For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
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The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
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During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
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The flow of genetic information in cells from DNA to mRNA to protein is described by the central dogma, which states that genes specify the sequence of mRNAs, which in turn specify the sequence of amino acids making up all proteins. The decoding of one molecule to another is performed by specific proteins and RNAs. Because the information stored in DNA is so central to cellular function, it makes intuitive sense that the cell would make mRNA copies of this information for protein synthesis...
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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...
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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...
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Updated: Feb 14, 2026

Separation of Single-stranded DNA, Double-stranded DNA and RNA from an Environmental Viral Community Using Hydroxyapatite Chromatography
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Fluorescent single-stranded DNA-binding protein from Plasmodium falciparum as a biosensor for single-stranded DNA.

Liisa T Chisty1, Daniela Quaglia1, Martin R Webb1

  • 1The Francis Crick Institute, London, United Kingdom.

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Researchers developed a new fluorescent biosensor using Plasmodium falciparum single-stranded DNA binding protein (PfSSB) for accurate measurement of ssDNA. This improved tool simplifies ssDNA quantitation in real-time cellular assays.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Single-stranded DNA (ssDNA) is transiently formed during crucial cellular processes like DNA replication and repair.
  • Measuring ssDNA formation is vital for understanding these fundamental biological mechanisms.
  • Existing biosensors, like DCC-EcSSB, faced limitations due to complex binding modes affecting ssDNA quantitation.

Purpose of the Study:

  • To develop an improved fluorescent biosensor for sensitive and accurate detection of ssDNA.
  • To utilize a novel scaffold, tetrameric SSB from Plasmodium falciparum, for enhanced biosensor performance.
  • To enable simple and reliable quantitation of ssDNA in various cellular contexts.

Main Methods:

  • Engineered a tetrameric SSB from Plasmodium falciparum, labeling each subunit with a diethylaminocoumarin fluorophore at a single surface site.
  • Characterized the fluorescent properties of the labeled SSB (DCC-PfSSB), observing a 20-fold fluorescence increase upon ssDNA binding.
  • Assessed the binding kinetics and affinity, determining a dissociation constant <5 pM for DCC-PfSSB.

Main Results:

  • The DCC-PfSSB biosensor exhibits a significant fluorescence enhancement upon binding to ssDNA, indicating its utility.
  • PfSSB demonstrates a consistent binding mode (65-70 bases per tetramer) across conditions, simplifying data interpretation.
  • The biosensor was successfully applied in real-time assays, exemplified by measuring AddAB helicase activity during double-stranded DNA unwinding.

Conclusions:

  • The novel DCC-PfSSB biosensor offers a significant improvement for ssDNA detection and quantitation.
  • Its simple binding mode and high affinity make it ideal for real-time monitoring of cellular processes involving ssDNA.
  • This tool facilitates a deeper understanding of DNA metabolism and related cellular functions.