Related Experiment Videos
Long-range structure of H-ras 1-selected transgenomes
W A Bickmore1, J C Maule, V van Heyningen
1MRC Human Genetics Unit, Western General Hospital, Edinburgh, Scotland.
Somatic Cell and Molecular Genetics
|May 1, 1989
Summary
Chromosome-mediated gene transfer (CMGT) can cause DNA rearrangements, unlike whole cell fusion. Understanding these molecular changes is crucial for using gene transfer techniques in genetic mapping and DNA probe isolation.
Area of Science:
- Somatic cell genetics
- Molecular biology
- Genomics
Background:
- Chromosome-mediated gene transfer (CMGT) and whole cell fusion are techniques used to create hybrid cells for genetic studies.
- Human-mouse hybrid cells with reduced chromosome 11 were derived by selecting for the H-ras 1 oncogene.
Purpose of the Study:
- To investigate molecular rearrangements during DNA transfer using CMGT and whole cell fusion.
- To assess the integrity of transferred DNA for applications in genetic mapping and DNA probe isolation.
Main Methods:
- Chromosome-mediated gene transfer (CMGT)
- Whole cell fusion
- Pulsed field gel electrophoresis (PFGE) to analyze DNA structure
Main Results:
- Whole cell fusion can transfer large DNA fragments (megabase pairs) intact.
- CMGT resulted in multiple DNA rearrangements, even when other analyses suggested structural integrity.
Conclusions:
- CMGT introduces significant DNA molecular scrambling, impacting its utility for precise genetic mapping.
- PFGE is essential for detecting subtle DNA rearrangements missed by other methods in CMGT-derived transgenomes.