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Identification and molecular cloning of Moloney mouse sarcoma virus-specific sequences from uninfected mouse cells
Abstract:
When uninfected mouse cell DNA is cleaved with restriction endonuclease EcoRI, a DNA fragment of 14.0 kilobases can be identified by hybridization to cloned DNA containing sarcoma specific sequences of Moloney mouse sarcoma virus (M-MSVsrc). The cellular DNA fragment contains the entire M-MSVsrc specific sequences. The 14.0-kilobase EcoRI DNA fragment was cloned in bacteriophage lambda. The sequence organization of a recombinant clone, lambda . MTX-1, was analyzed by restriction endonuclease mapping, nuclease S1 mapping, and electron microscopy. The results indicate that lambda . MTX-1 contains an uninterrupted stretch of 1.0 kilobase similar to that found in the M-MSV genome.
Insights
Researchers identified a 14.0-kilobase DNA fragment containing Moloney mouse sarcoma virus (M-MSVsrc) sequences. This fragment was cloned and analyzed, revealing an uninterrupted 1.0 kilobase sequence similar to the M-MSV genome.
Area of Science:
- Molecular Biology
- Virology
- Genetics
Background:
- The Moloney mouse sarcoma virus (M-MSV) is a retrovirus known to carry sarcoma-specific sequences (M-MSVsrc).
- Understanding the integration and organization of viral genetic material within host DNA is crucial for studying viral pathogenesis and oncogenesis.
Purpose of the Study:
- To characterize the cellular DNA fragment containing the M-MSVsrc specific sequences.
- To clone and analyze the sequence organization of this cellular DNA fragment.
Main Methods:
- Restriction endonuclease digestion (EcoRI) of mouse cell DNA.
- Hybridization using cloned M-MSVsrc DNA.
- Cloning of the 14.0-kilobase fragment into bacteriophage lambda.
- Sequence analysis of the recombinant clone (lambda . MTX-1) using restriction endonuclease mapping, nuclease S1 mapping, and electron microscopy.
Main Results:
- A 14.0-kilobase EcoRI-cleaved DNA fragment was isolated from uninfected mouse cells.
- This fragment hybridized to M-MSVsrc sequences and contained the entire M-MSVsrc specific sequences.
- The recombinant clone lambda . MTX-1 contained an uninterrupted 1.0 kilobase sequence homologous to the M-MSV genome.
Conclusions:
- The cellular DNA contains a complete M-MSVsrc sequence integrated within a 14.0-kilobase fragment.
- The cloned M-MSVsrc sequence in lambda . MTX-1 is structurally similar to that in the M-MSV genome, suggesting conservation of viral genetic elements.