The effect of monofunctional or difunctional platinum adducts and of various other associated DNA damage on the expression of transfected DNA in mammalian cell lines sensitive or resistant to difunctional agents

1987 ◽  
Vol 908 (3) ◽  
pp. 214-223 ◽  
Author(s):  
R.J. Knox ◽  
D.A. Lydall ◽  
F. Friedlos ◽  
C. Basham ◽  
J.J. Roberts
Keyword(s):  
Dna Damage ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Mammalian Cell Lines

Comparative analysis of micronuclei and DNA damage induced by Ochratoxin A in two mammalian cell lines

2011 ◽  
Vol 723 (1) ◽  
pp. 58-64 ◽  
Author(s):  
Rahat Ali ◽  
Roberta A. Mittelstaedt ◽  
Joseph G. Shaddock ◽  
Wei Ding ◽  
Javed A. Bhalli ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comparative Analysis ◽  
Ochratoxin A ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Mammalian Cell Lines

DNA damage in mammalian cell lines with different antioxidant levels and DNA repair capacities

1992 ◽  
pp. 247-250
Author(s):  
Bernadette M. Hannigan ◽  
Shirley-Ann M. Richardson ◽  
P. Gerald McKenna
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Mammalian Cell Lines

Ochratoxin A: induction of (oxidative) DNA damage, cytotoxicity and apoptosis in mammalian cell lines and primary cells

Toxicology ◽  
2005 ◽  
Vol 206 (3) ◽  
pp. 413-425 ◽  
Author(s):  
H KAMP ◽  
G EISENBRAND ◽  
J SCHLATTER ◽  
K WURTH ◽  
C JANZOWSKI
Keyword(s):  
Dna Damage ◽  
Ochratoxin A ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Oxidative Dna Damage ◽  
Primary Cells ◽  
Mammalian Cell Lines

scholarly journals Structure-guided selection of puromycin N-acetyltransferase mutants with enhanced selection stringency for deriving mammalian cell lines expressing recombinant proteins

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alessandro T. Caputo ◽  
Oliver M. Eder ◽  
Hana Bereznakova ◽  
Heleen Pothuis ◽  
Albert Ardevol ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mammalian Cell ◽  
Recombinant Proteins ◽  
Cultured Cells ◽  
Hek293 Cells ◽  
Reaction Products ◽  
Enzyme Form ◽  
X Ray Crystallography ◽  
Mammalian Cell Lines ◽  
Apparent Loss

AbstractPuromycin and the Streptomyces alboniger-derived puromycin N-acetyltransferase (PAC) enzyme form a commonly used system for selecting stably transfected cultured cells. The crystal structure of PAC has been solved using X-ray crystallography, revealing it to be a member of the GCN5-related N-acetyltransferase (GNAT) family of acetyltransferases. Based on structures in complex with acetyl-CoA or the reaction products CoA and acetylated puromycin, four classes of mutations in and around the catalytic site were designed and tested for activity. Single-residue mutations were identified that displayed a range of enzymatic activities, from complete ablation to enhanced activity relative to wild-type (WT) PAC. Cell pools of stably transfected HEK293 cells derived using two PAC mutants with attenuated activity, Y30F and A142D, were found to secrete up to three-fold higher levels of a soluble, recombinant target protein than corresponding pools derived with the WT enzyme. A third mutant, Y171F, appeared to stabilise the intracellular turnover of PAC, resulting in an apparent loss of selection stringency. Our results indicate that the structure-guided manipulation of PAC function can be utilised to enhance selection stringency for the derivation of mammalian cell lines secreting elevated levels of recombinant proteins.

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