Related Experiment Video

Updated: Aug 14, 2026

High Sensitivity 5-hydroxymethylcytosine Detection in Balb/C Brain Tissue
07:19

High Sensitivity 5-hydroxymethylcytosine Detection in Balb/C Brain Tissue

Published on: February 2, 2011

MspI, an isoschizomer of hpaII which cleaves both unmethylated and methylated hpaII sites

C Waalwijk, R A Flavell

    Nucleic Acids Research
    |September 1, 1978
    PubMed

    Insights

    Methylation at the internal C residue of CCGG sites blocks DNA cleavage by HpaII and HapII. However, the isoschizomer MspI cleaves methylated DNA, distinguishing between methylated and unmethylated sequences.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Epigenetics

    Background:

    • Restriction endonucleases like HpaII and HapII recognize the CCGG sequence.
    • DNA methylation at the internal cytosine residue (C) within the CCGG sequence inhibits cleavage by HpaII and HapII.
    • This methylation pattern is prevalent in mammalian sperm DNA.

    Purpose of the Study:

    • To investigate the differential cleavage activity of restriction enzymes HpaII, HapII, and MspI on methylated and unmethylated DNA.
    • To explore the utility of MspI in distinguishing between methylated and unmethylated CCGG sites.

    Main Methods:

    • Comparative DNA digestion assays using restriction endonucleases HpaII, HapII, and MspI.
    • Analysis of DNA cleavage patterns in bacterial DNA (Haemophilus parainfluenzae, Haemophilus aphrophilus) and mammalian sperm DNA.
    • Determination of average DNA fragment sizes after enzymatic digestion.

    Main Results:

    • HpaII and HapII failed to cleave DNA containing 5-methylcytosine at the internal C of CCGG sites.
    • MspI, an isoschizomer of HpaII, efficiently cleaved DNA irrespective of methylation status at the internal C.
    • MspI demonstrated differential cleavage of mammalian sperm DNA (average size ~5 kb) compared to HpaII/HapII (average size ~40 kb), indicating extensive internal C methylation.

    Conclusions:

    • MspI can effectively cleave DNA at CCGG sites regardless of internal cytosine methylation.
    • This differential activity allows MspI to serve as a valuable tool for detecting and analyzing DNA methylation patterns.
    • The study highlights the prevalence of methylation at HpaII/HapII sites in mammalian sperm DNA.

    More Related Videos

    LINE-1 Methylation Analysis in Mesenchymal Stem Cells Treated with Osteosarcoma-Derived Extracellular Vesicles12:18

    LINE-1 Methylation Analysis in Mesenchymal Stem Cells Treated with Osteosarcoma-Derived Extracellular Vesicles

    Published on: February 1, 2020

    Identification of Modified Histones as Binding Substrates of Human Spindlin Family Member 4 (SPIN4) by Peptide Arrays and Native Nucleosome Pulldown09:34

    Identification of Modified Histones as Binding Substrates of Human Spindlin Family Member 4 (SPIN4) by Peptide Arrays and Native Nucleosome Pulldown

    Published on: March 27, 2026

    Related Concept Videos

    Conservative Site-specific Recombination and Phase Variation02:53

    Conservative Site-specific Recombination and Phase Variation

    Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
    The recognition sites for Cre recombinase called LoxP...
    piRNA - Piwi-interacting RNAs02:57

    piRNA - Piwi-interacting RNAs

    PIWI-interacting RNAs, or piRNAs, are the most abundant short non-coding RNAs. More than 20,000 genes have been found in humans that code for piRNAs while only 2000 genes have been found for miRNAs. piRNAs can act at the transcriptional and post-transcriptional levels and have a vital role in silencing transposable elements present in germ cells. They are also involved in epigenetic silencing and activation. Previously, they were thought to function only in germ cells but new evidence suggests...
    Single-Strand DNA Binding Proteins01:03

    Single-Strand DNA Binding Proteins

    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...