Document Detail


Chromosome-mediated gene transfer results in two classes of unstable transformants.
MedLine Citation:
PMID:  6932038     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The human thymidine kinase gene has been transferred from HeLa S3 cells to mouse LM(TK-) cells via isolated metaphase chromosomes. Efficient transfer of the thymidine kinase gene (1.8 X 10(-5) colonies per recipient cell) was obtained when the donor chromosomes were precipitated with calcium phosphate and the recipient cells were treated with 10% (vol/vol) dimethyl sulfoxide. Thirty-five independent cell lines were analyzed in detail. Cytologically detectable donor chromosome fragments were observed in 14% of the cell lines. Many of the transformed cell lines were also found to express the human genes for galactokinase (23% of the transformed cell lines) and procollagen type I (69% of the transformed cell lines), which are syntenic to thymidine kinase on human chromosome 17. On the basis of stability analyses, three classes of transformed cell lines were defined and characterized. One class of transformants was stable, showing no loss of the transferred phenotype in the absence of selection. A second group of transformants was unstable, losing the thymidine kinase phenotype at a rate of 1.5-2.5% per day. This group of transformants was found to possess large donor chromosome fragments (macrotransgenomes) and relatively low levels of donor gene activity. The third group of transformants lost the thymidine kinase phenotype rapidly, at a rate of 6-10% per day. These cell lines contained small, cytologically undetectable transgenomes (microtransgenomes) and overexpressed the transferred thymidine kinase gene.
Authors:
L A Klobutcher; C L Miller; F H Ruddle
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  77     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1980 Jun 
Date Detail:
Created Date:  1980-11-25     Completed Date:  1980-11-25     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  3610-4     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Chromosomes, Human, 16-18*
Galactokinase / genetics
Humans
Hybrid Cells / classification,  enzymology
Karyotyping
Linkage (Genetics)
Mice
Phenotype
Procollagen / genetics
Selection, Genetic
Thymidine Kinase / analysis,  genetics*
Time Factors
Transformation, Genetic*
Chemical
Reg. No./Substance:
0/Procollagen; EC 2.7.1.21/Thymidine Kinase; EC 2.7.1.6/Galactokinase
Comments/Corrections

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