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  4. Use of fluorescence in situ hybridization to detect and monitor transfected and amplified sequences in recombinant CHO cells
 
research article

Use of fluorescence in situ hybridization to detect and monitor transfected and amplified sequences in recombinant CHO cells

Pallavicini, M. G.
•
DeTeresa, P. S.
•
Wurm, F. M.  
1989
Developments in biological standardization

Non-radioactive fluorescence in situ hybridization was used to detect transfected and amplified sequences in recombinant CHO cells. The cell lines used in this study are based on DHFR-deficient CHO-DUKX cells. The recombinant CHO cells contain and express, as verified by Southern and Northern experiments, multiple copies of a constitutive DHFR expression vector, as well as an inducible Drosophila HSP 70 promoter-mouse c-myc construction. In order to localize and monitor the chromosomal location of transfected and amplified DHFR and c-myc sequences, biotinylated DNA probes were hybridized to metaphase preparations of several cell lines. The resulting hybrids were detected using fluorescein-linked avidin. The fluorescence signal was amplified using a biotinylated anti-avidin antibody. The number of c-myc and DHFR integration sites per metaphase, their distribution in cell populations growing at various methotrexate levels, the sizes of the amplified sequences, as well as the number of chromosomal rearrangements were measured. The results of this study will be presented and the usefulness of this method as a general tool for rapid characterization and monitoring of recombinant cell lines will be discussed.

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Type
research article
PubMed ID

2668071

Author(s)
Pallavicini, M. G.
DeTeresa, P. S.
Wurm, F. M.  
Date Issued

1989

Published in
Developments in biological standardization
Volume

70

Start page

165

End page

72

Subjects

Animals

•

Cell Line

•

Cricetinae

•

Cricetulus

•

DNA Probes

•

DNA

•

Recombinant/*analysis

•

Female

•

*Gene Amplification

•

Guinea Pigs

•

Microscopy

•

Fluorescence

•

Nucleic Acid Hybridization

•

Ovary

•

Recombinant Proteins/*biosynthesis

•

Transfection/*methods

Note

Biomedical Sciences Division, Lawrence Livermore National Laboratory, Livermore, CA 94550.

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

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Available on Infoscience
June 5, 2007
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/7584
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