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大量含有大肠杆菌乳糖操纵子控制元件的限制性片段的制备与表征。

Preparation and characterization of large amounts of restriction fragments containing the E. coli lac control elements.

作者信息

Hardies S C, Wells R D

出版信息

Gene. 1979 Sep;7(1):1-14. doi: 10.1016/0378-1119(79)90039-8.

Abstract

Large quantities of pure DNA fragments (789, 203 and 95 bp in length) containing the Escherichia coli lac controlling elements (operator, promoter, CRP binding site) were prepared from appropriate recombinant plasmids. High pressure liquid chromatography on RPC-5 or preparative sucrose gradient centrifugation was used to fractionate the pVH51 vector from the inserts. The fragments had few, if any, nicks or depurinated sites, and the majority of the fragment ends were intact. Absorbance-temperature profiles on the fragments showed multiphasic transitions.

摘要

从合适的重组质粒中制备了大量含有大肠杆菌乳糖操纵元件(操纵基因、启动子、CRP结合位点)的纯DNA片段(长度分别为789、203和95碱基对)。使用RPC - 5上的高压液相色谱或制备性蔗糖梯度离心法将pVH51载体与插入片段分离。这些片段即使有切口或脱嘌呤位点也很少,并且大多数片段末端是完整的。片段的吸光度 - 温度曲线显示出多相转变。

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