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[在大肠杆菌中克隆叶绿体DNA。I. 含有豌豆叶绿体DNA片段的重组质粒的构建与筛选]

[Cloning chloroplast DNA in escherichia coli. I. Construction and selection of recombinant plasmids containing fragments of pea chloroplast DNA].

作者信息

Andrianov V M, Zemlianaia N Iu, Karimov M, Amerkhanova M B, Vinetskiĭ Iu Ts

出版信息

Genetika. 1978 Sep;14(9):1503-12.

PMID:363514
Abstract

Fragments produced by digestion of Pisum sativum chloroplast DNA with EcoRI were examined by agarose gel electrophoresis. These EcoRI-fragments were joined in vitro to Apr-ColE1 RSF2124 plasmid and cloned in Escherichia coli. Methods of molecular cloning of plasmid chimeras by success gradient centrifugation and repeated transformation and selection of recombinant plasmids using mytomicin C were used for cloning hybrid plasmids with various EcoRI fragments of pea chloroplast DNA has been obtained.

摘要

用EcoRI消化豌豆叶绿体DNA产生的片段通过琼脂糖凝胶电泳进行检测。这些EcoRI片段在体外与Apr-ColE1 RSF2124质粒连接,并克隆到大肠杆菌中。通过连续梯度离心克隆质粒嵌合体以及使用丝裂霉素C对重组质粒进行重复转化和筛选的方法,已获得了含有豌豆叶绿体DNA各种EcoRI片段的杂交质粒。

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