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Example sentences for: tkt
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Characterization of tkT lines
We have previously characterized clonal mouse astrocyte cell lines, immortalized by transduction of SV40 large T antigen expressed from the MoMuLV LTR [ 6 13 ] . Here, we compare variation in Tag levels in 3 of these lines and 3 additional lines (termed tkT lines), immortalized with Moloney Sarcoma Virus vectors expressing Tag from an internal herpes thymidine kinase (tk) promoter.
The remaining two essential gene products were in the pentosephosphate pathway ( tktAB, rpiAB ). The tktAB and rpiAB gene products were required for the synthesis of erythrose 4-phosphate in all phases (aromatic amino acid supplement required for the cultivation of tkt - E. coli mutant strains [ 27]).
Cortical astrocytes were harvested from neonatal C57BL/6 mice and infected with a replication-defective retrovirus encoding Tag and neomycin resistance as previously described [ 13 ] . The transferred viruses were SV40-6 [ 5 ] , in which wild type Tag is expressed from the M-MuLV LTR promoter; SV(X)T K1, which is constructed the same as SV40-6 except that the Tag is mutant [ 38 ] and defective for binding the retinoblastoma protein family [ 39 ] ; Linker-tkT, in which the cDNA for Tag is expressed from the herpes simplex 1 tk promoter [ 11 ] . The differentiation and transformation phenotypes of the cell lines utilizing the M-MuLV LTR promoter have been described [ 6 13 ] . The Linker-tkT lines (tkT) were either isolated as clones after G418 selection (as previously described) [ 6 ] or frozen immediately after selection (pooled clones) and clonally selected at a later time.
To test this, the cell line in which Tag control was most sensitive to density, P0-3D tkT#5, was grown for 3 days starting at an intermediate density of 10 6cells per 10 cm plate with varying concentrations of serum ranging from 2.5-10%.
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