Specific inhibition of DNA biosynthesis induced by 3'-amino-2',3'-dideoxycytidine.
Specific inhibition of DNA biosynthesis induced by 3'-amino-2',3'-dideoxycytidine.
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特异性抑制 3-氨基-2,3-二脱氧胞苷诱导的 DNA 生物合成。
DOI：
10.1021/bi00424a022
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发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
Lin,TS
中科院分区：
文献类型：
--
作者：
Mancini,WR;Williams,MS;Lin,TS
材料与方法已经报道了3 '-NH 2-dCyd的合成（Lin & Mancini，1983; Lin等人，1983年）。[2，8- 3 H]脱氧腺苷（25 Ci/mmol）、[甲基-14 H]胸苷（70 Ci/mmol）、[2- 14 C]胸苷（56 mCi/mmol）和[5，6- 3 H]-nra-C（26 Ci/mmol）购自Moravek Biochemicals，Inc.。(Brea，CA）。[5- 3 H]脱氧胞苷S ′-三磷酸（30 Ci/mmol）购自ICN Chemical & Radiochemical Division（Irvine，CA），而无载体的[32 P]原磷酸由新英格兰核能公司（Boston，MA）提供。链霉蛋白酶（76600蛋白水解单位/g）获自Calbiochem-Behring（San Diego，CA），而脱氧核糖核酸酶I（3300 Kunitz单位/mg）、核糖核酸酶A（79 Kunitz单位/mg）、核酸酶P2（1600单位/mg）和核苷-S ′-二磷酸激酶（1000单位/mg）获自Sigma Chemical Co. (St.密苏里州路易斯）。核苷酸单磷酸激酶（0.5单位/mg）购自Boehringer曼海姆Biochemicals（Indianapolis，IN），Dowex AG 50 W-X4（氢形式）购自Bio-Rad Laboratories（里士满，CA）。L1210白血病细胞在补充有10%马血清的Fischer培养基（Grand Island Biological Co.，Grand Island，NY）。本研究仅使用处于生长基本期的细胞。通过生物培养方法发现细胞无支原体污染（Barile，1973）。流式细胞仪为了减少样品的可变性，
Materials and MethodsMaterials. Thesynthesis of 3'-NH2-dCyd has been reported (Lin & Mancini, 1983; Lin et al., 1983).[2, 8-3H] Deoxy-adenosine (25 Ci/mmol),[methyl-^ H] thymidine (70 Ci/mmol),[2-14C] thymidine (56 mCi/mmol), and [5, 6-3H]-nra-C (26 Ci/mmol) were purchased from Moravek Biochemicals, Inc.(Brea, CA).[5-3H] DeoxycytidineS'-triphosphate (30 Ci/mmol) was obtained from ICN Chemical & Radiochemical Division (Irvine, CA) while carrier-free [32P] ortho-phosphate was supplied by New England Nuclear (Boston, MA). Pronase (76 600 proteolytic units/g) was obtained from Calbiochem-Behring (San Diego, CA) while deoxyribonuclease I (3300 Kunitz units/mg), ribonuclease A (79 Kunitz un-its/mg), nuclease P2 (1600 units/mg), and nucleoside-S'-di-phosphate kinase (1000 units/mg) were obtained from Sigma Chemical Co.(St. Louis, MO). The nucleosidemonophosphate kinase (0.5 unit/mg) was purchased from Boehringer Mannheim Biochemicals (Indianapolis, IN) and Dowex AG 50W-X4 (hydrogen form) from Bio-Rad Laboratories (Richmond, CA).Cells. L1210 leukemia cells were grown in Fischer’s me-dium supplemented with 10% horse serum(Grand Island Biological Co., Grand Island, NY). Only cells in theexpo-nential phase of growth were used in this investigation. Cells were found to be free of mycoplasma contamination by a biological culture method (Barile, 1973). Flow Cytometry. In an effort to reduce sample variability,