DNA‐mediated gene transfer as an indicator of DNA damage and its repair by recipient cells

FEBS Letters

Ireland, Christine M.¹  Stewart, Bernard W.¹ 

[1] Children's Leukaemia and Cancer Research Unit, Prince of Wales Children's Hospital, Randwick 2031, New South Wales, Australia

关键词: DNA repair;    Transfection;    Thymidine kinase gene;    Methylmethanesulfonate;    N-Methyl-N-nitrosourea;    ara-C;    1-β-arabinofuranosyl cytosine;    ddT;    dideoxythymidine;    MMS;    methyl methanesulphonate;    MNU;    N-methyl-N-nitrosourea;    TK;    thymidine kinase;   

DOI  :  10.1016/0014-5793(87)81580-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Preparations of plasmid containing the thymidine kinase gene (pHSV106) were treated with the alkylating agents methyl methanesulphonate or N-methyl-N-nitrosourea prior to transfection into thymidine kinase-deficient mouse L-cells using the DNA-calcium phosphate co-precipitation technique. Relative to transfection with unmodified plasmid, a reduced transformation efficiency was observed using alkylation-damaged plasmid, N-methyl-N-nitrosourea causing the greatest inhibition. Treatment of recipient cells with arabinosyl cytosine or dideoxythymidine during the expression period following transfection by the ‘damaged’ plasmid reduced transformation efficiency, suggesting that DNA repair 4–6 h post-transfection was required for gene expression.

【 授权许可】

Unknown