A sectored colony assay for monitoring mutagenesis by specific carcinogen-DNA adducts in Escherichia coli.

A sectored colony assay for monitoring mutagenesis by specific carcinogen-DNA adducts in Escherichia coli.
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用于监测大肠杆菌中特定致癌物 DNA 加合物诱变的扇形菌落测定。

DOI:
10.1021/bi00114a014
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发表时间:
1991
期刊:
影响因子:
2.9
通讯作者:
Moschel,RC
Moschel,RC
中科院分区:
生物学3区
文献类型:
--
作者:
Pauly,GT;Hughes,SH;Moschel,RC

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材料与方法酶RspMI、NdeI、Xbal和T4多核苷酸激酶购自新英格兰Biolabs,Beverly,MA。E.大肠杆菌C1 - 5-烷基鸟嘌呤-DNA烷基转移酶(ADA蛋白)获自Applied Genetics,Inc.,纽约州弗里波特市。T4 DNA连接酶、5-溴-4-氯-3-吲哚基/SD-半乳糖苷和异丙基/SD-硫代-半乳糖苷(IPTG)来自United States Biochemicals,Cleveland,OH。N-甲基-N,N-硝基-N-亚硝基胍(MNNG)来自Sigma Chemical Co.,密苏里州圣路易斯其它试剂和溶剂来自Aldrich Chemical Co.,股份有限公司、密尔沃基,WI,American Burdick和杰克逊,Muskegon,MI,应用生物系统公司,Foster City,CA或Pharmacia LKB Biotechnology,Inc.,新泽西州,皮斯卡特维。寡核苷酸合成和纯化。这些研究中使用的16个碱基序列(图1)在Applied Biosystems,Inc. 380 B型DNA合成仪。
Materials and MethodsMaterials. The enzymes RspMI, NdeI, Xbal, and T4 po-lynucleotide kinase were purchased from New England Biolabs, Beverly, MA. E. coli C^-alkylguanine-DNA alkyl-transferase (the ada protein) was obtained from Applied Genetics, Inc., Freeport, NY. T4 DNA ligase, 5-bromo-4-chloro-3-indolyl/SD-galactoside, and isopropyl/SD-thio-galactoside (IPTG) were from United States Biochemicals, Cleveland, OH. TV-Methyl-jV'-nitro-iV-nitrosoguanidine (MNNG) was from Sigma Chemical Co., St. Louis, MO. Other reagents and solvents were from Aldrich Chemical Co., Inc., Milwaukee, WI, American Burdick and Jackson, Muskegon, MI, Applied Biosystems, Inc., Foster City, CA, or Pharmacia LKB Biotechnology, Inc., Piscataway, NJ. Oligonucleotide Synthesis and Purification. The 16-base sequences used in these studies (Figure 1) were synthesized on an Applied Biosystems, Inc., Model 380B DNA synthesizer.
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DOI: --
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发表时间: 1990-11-01
期刊: MUTATION RESEARCH
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