scholarly journals Procaryotic genomic DNA inhibits mammalian cell transformation.

1983 ◽  
Vol 3 (5) ◽  
pp. 956-959 ◽  
Author(s):  
J I Yoder ◽  
A T Ganesan
Keyword(s):  
Thymidine Kinase ◽  
Mammalian Cell ◽  
Genomic Dna ◽  
Cell Transformation ◽  
Phage Dna ◽  
Mouse Cells

Ltk- mouse cells were transformed to thymidine kinase prototrophy in the presence of carrier DNAs isolated from different organisms. Procaryotic genomic and phage DNA was consistently less effective as a carrier than was eucaryotic DNA. Mixing experiments indicate that DNA of procaryotic origin inhibits mammalian cell transformation.

Procaryotic genomic DNA inhibits mammalian cell transformation

1983 ◽  
Vol 3 (5) ◽  
pp. 956-959
Author(s):  
J I Yoder ◽  
A T Ganesan
Keyword(s):  
Thymidine Kinase ◽  
Mammalian Cell ◽  
Genomic Dna ◽  
Cell Transformation ◽  
Phage Dna ◽  
Mouse Cells

Ltk- mouse cells were transformed to thymidine kinase prototrophy in the presence of carrier DNAs isolated from different organisms. Procaryotic genomic and phage DNA was consistently less effective as a carrier than was eucaryotic DNA. Mixing experiments indicate that DNA of procaryotic origin inhibits mammalian cell transformation.

Correction of Genetic Defects in Mammalian Cells by the Input of Small Amounts of Foreign Genetic Material

1973 ◽  
Vol 13 (3) ◽  
pp. 841-861
Author(s):  
YVONNE L. BOYD ◽  
H. HARRIS
Keyword(s):  
Thymidine Kinase ◽  
Mammalian Cells ◽  
Hybrid Cell ◽  
Genetic Material ◽  
Chinese Hamster ◽  
Heat Sensitivity ◽  
Chinese Hamster Cells ◽  
Inosinic Acid ◽  
Mouse Cells

Chinese hamster cells lacking inosinic acid pyrophosphorylase and mouse cells lacking thymidine kinase were fused with chick erythrocytes. The resultant heterokaryons were cultivated in a selective medium in which possession of these enzymes was essential for cell survival and growth. Clones of cells able to grow in this medium were isolated and studied. A detailed karyological analysis of these clones failed to reveal any chick chromosomes; nor could any chick-specific antigens be detected on the surface of the cells. Nonetheless, clones arising from the fusion of chick erythrocytes with Chinese hamster cells were shown to possess an inosinic acid pyrophosphorylase which had the electrophoretic characteristics of chick inosinic acid pyrophosphorylase. However, the clones arising from the fusion of the chick erythrocytes with the mouse cells had a thymidine kinase with the electrophoretic mobility and heat sensitivity of murine, not chick, thymidine kinase. Both types of hybrid cell have now been cultivated in vitro for 18 months without the loss of thymidine kinase or inosinic acid pyrophosphorylase activity.

A stable bovine papillomavirus hybrid plasmid that expresses a dominant selective trait

1983 ◽  
Vol 3 (11) ◽  
pp. 2110-2115
Author(s):  
M F Law ◽  
J C Byrne ◽  
P M Howley
Keyword(s):  
Resistance Gene ◽  
Mammalian Cell ◽  
Morphological Characteristics ◽  
Hybrid Plasmid ◽  
Transcriptional Unit ◽  
Bovine Papillomavirus ◽  
G418 Resistance ◽  
Extrachromosomal Element ◽  
Mouse Cells ◽  
Neomycin Resistance

We describe a bovine papillomavirus hybrid plasmid containing the neomycin resistance gene from Tn5 inserted into a mammalian cell transcriptional unit. This plasmid is maintained as a stable extrachromosomal element (20 to 100 copies per diploid genome) in mouse cells selected either for the transformed phenotype or for resistance to the aminoglycoside G418. Cells selected for G418 resistance initially display a flat, nontransformed phenotype before exhibiting the gross morphological characteristics of transformation. The delay in the appearance of the transformed phenotype indicated that some intracellular event or series of events subsequent to the establishment of transcriptionally active bovine papillomavirus 1 hybrid plasmid is required for the manifestation of the transformed phenotype.

Isolation and preliminary characterization of the Chinese hamster thymidine kinase gene

1983 ◽  
Vol 3 (10) ◽  
pp. 1815-1823
Author(s):  
J A Lewis ◽  
K Shimizu ◽  
D Zipser
Keyword(s):  
Thymidine Kinase ◽  
Genomic Dna ◽  
Chinese Hamster ◽  
Growth Conditions ◽  
Phage Library ◽  
Hybridization Technique ◽  
Kinase Gene ◽  
Recombinant Phage ◽  
Hybridization Probes

The Chinese hamster thymidine kinase (TK) gene has been isolated from a recombinant phage library constructed with genomic DNA from mouse Ltk- cells transformed to Tk+ by transfection with Chinese hamster genomic DNA. The phage library was screened by the Benton-Davis plaque hybridization technique, using as probes, subclones of recombinant phage that were isolated from mouse Ltk+ transformants by the tRNA suppressor rescue method. The Chinese hamster TK gene is contained within 13.2 kilobases of genomic DNA in the isolate designated lambda 34S4. This gene, defined by restriction enzyme sensitivity experiments, homology studies with the chicken TK gene, and mRNA blotting experiments, may extend over 8.5 kilobases. Subclones of the lambda 34S4 isolate used as hybridization probes identified a 1,400-nucleotide polyadenylated RNA as the hamster TK mRNA. The abundance of this mRNA varies dramatically in Chinese hamster cells cultured under various growth conditions, providing direct evidence that the growth dependence of TK activity may be regulated in an important way at the level of cytoplasmic TK mRNA.

Transformation methods for halophilic archaebacteria

1989 ◽  
Vol 35 (1) ◽  
pp. 148-152 ◽  
Author(s):  
Steven W. Cline ◽  
Wan L. Lam ◽  
Robert L. Charlebois ◽  
Leonard C. Schalkwyk ◽  
W. Ford Doolittle
Keyword(s):  
Molecular Weight ◽  
Plasmid Dna ◽  
Genomic Dna ◽  
Halobacterium Halobium ◽  
Dna Transformation ◽  
Dna Transfection ◽  
Phage Dna ◽  
Halophilic Archaebacteria ◽  
Transformation Methods ◽  
Halobacterium Volcanii

We present a practical description of polyethylene glycol mediated spheroplast transformation of Halobacterium halobium and Halobacterium volcanii. This method has been applied to phage DNA transfection, plasmid DNA transformation, and transformation with linear fragments of high molecular weight genomic DNA. Efficient spheroplast regeneration allows uncomplicated recovery of transformed progeny. Transformations can be performed equally well using fresh or frozen cell preparations. These methods should find application in molecular cloning, genetic fine mapping, and strain construction.Key words: archaebacteria, Halobacterium, transformation methods, spheroplast.

Human Thymidine Kinase Gene Locus: Assignment to Chromosome 17 in a Hybrid of Man and Mouse Cells

Science ◽  
1971 ◽  
Vol 173 (3993) ◽  
pp. 244-245 ◽  
Author(s):  
O. J. Miller ◽  
P. W. Allderdice ◽  
D. A. Miller ◽  
W. R. Breg ◽  
B. R. Migeon
Keyword(s):  
Thymidine Kinase ◽  
Gene Locus ◽  
Chromosome 17 ◽  
Thymidine Kinase Gene ◽  
Kinase Gene ◽  
Mouse Cells
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