Construction and characterization of a site-directed CC-1065-N3-adenine adduct within a 117 base pair DNA restriction fragment.

Construction and characterization of a site-directed CC-1065-N3-adenine adduct within a 117 base pair DNA restriction fragment.
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117 碱基对 DNA 限制性片段内定点 CC-1065-N3-腺嘌呤加合物的构建和表征。

DOI:
10.1021/bi00374a016
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发表时间:
1986
期刊:
影响因子:
2.9
通讯作者:
Hurley,LH
Hurley,LH
中科院分区:
生物学3区
文献类型:
--
作者:
Needham-VanDevanter,DR;Hurley,LH

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材料和方法材料CC-1065购自密歇根州Kalamazoo的Upjohn Co.,电泳试剂[丙烯酰胺、TEMED、过硫酸铵和丙烯酰胺]来自Bio-Rad。BAP、dNTPs和溶菌酶来自Sigma。所有其他酶[限制性内切酶、T4-PNK和DNA聚合酶I(大片段)]均来自New EnglandBiolabs。M13mpl载体来自明尼苏达大学J. Messing教授。MPE是加州理工学院P. Dervan教授的慷慨馈赠。M13mpl RF DNA从感染的大肠杆菌JM101细胞中用CsCl2超离心分离,方法如Messing(1983)所述。[t-32P] ATP来源于ICN。x光胶片、强化屏和显影剂都来自柯达。
Materials and MethodsMaterials CC-1065 was obtained from the Upjohn Co., Kalamazoo, MI. Electrophoretic reagents [acrylamide, TEMED, ammo-nium persulfate, and bis (acrylamide)] were from Bio-Rad. BAP, dNTPs, and lysozyme were from Sigma. All other enzymes [restriction endonucleases, T4-PNK, and DNA polymerase I (large fragment)] were from New EnglandBiolabs. M13mpl vector was obtained from Professor J. Messing, University of Minnesota. MPE was a generous gift of Pro-fessor P. Dervan, California Institute of Technology. M13mpl RF DNA was isolated from infected Escherichia coli JM101 cells by CsCl2 ultracentrifugation as described by Messing (1983).[t-32P] ATP was from ICN. X-ray film, intensifying screen, and developing chemicals were from Kodak.
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