Directed formation of deletions and duplications using Mud(Ap, lac).
Directed formation of deletions and duplications using Mud(Ap, lac).
复制标题
使用 Mud(Ap, lac) 定向形成缺失和重复。
DOI：
10.1093/genetics/109.2.263
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发表时间：
1985
期刊：
影响因子：
3.3
通讯作者：
Roth,JR
中科院分区：
文献类型：
--
作者：
Hughes,KT;Roth,JR
一种用于分离染色体缺失和带有预定终点的重复的遗传程序。这些重排是利用两个菌株上生长的P22转导噬菌体裂解物的混合物在转导杂交中产生的，每个菌株都带有泥-内酰胺插入。通过插入MUD_1-8(一种转座-缺陷型泥噬菌体)，证明了在组操纵子中形成了复制和缺失。这项技术还被用来在白细胞合成操纵子中的MUD_1-8插入之间和在T_P中的MUD插入之间进行更大的染色体复制。提供的遗传证据有力地表明，通过P22介导的杂交来遗传单个粘液噬菌体需要两个转导片段；每个片段携带部分粘液噬菌体。这两个片段必须参与三次重组交换；一次交换加入供体MudFrages，两个复合片段与受体染色体之间发生两次交换，一次在完整的供体Mudement的两侧。由于复制只发生在染色体上相同方向的MUDI插入之间，复制形成方法提供了一种简单的方法来确定MUD1-8在染色体上的方向，从而确定MUDIS插入的基因的转录方向。用该方法构建了MUD1-8噬菌体与Casadaban‘s蛋白融合载体Mud2之间的重组体，从而分离出Mud2-8，一种含有Mud2蛋白融合能力和MUD1-8转座缺陷功能的Mud2-8。
A genetic procedure is directed for the isolation of chromosomal deletions and duplications with predetermined endpoints. These rearrangements are generated in transduction crosses using a mixture of P22-transducing phage lysates grown on two strains, each carrying a Mud-lacinsertion. The formation of duplications and deletions was demonstrated in thehisoperon using insertions of Mud1-8(a transposition-defectiveMud-lacphage). This technique was also used to make larger chromosomal duplications between Mud1-8insertions in thethrandleubiosynthetic operons and between Mudinsertions in thethrandpyrBoperons. Genetic evidence is presented that strongly suggests that inheritance of a single Mudprophage by P22-mediated crosses requires two transduced fragments; each carrying part of the Mudprophage. The two fragments must be involved in three recombinational exchanges; one exchange joins the donor Mudfragments and two exchanges occur between the composite fragment and the recipient chromosome, one on either side of the complete donor Mudelement. Since duplications only occur between Mudinsertions in the same orientation on the chromosome, the method of duplication formation provides a simple means of determining the orientation of Mud1-8on the chromosome and, therefore, the direction of transcription of the gene into which Mudis inserted. This method was also used to construct recombinants between a Mud1-8prophage and Casadaban's protein fusion vector Mud2 and, thereby, isolate Mud2-8, a Mudderivative containing the protein fusion ability of Mud2 and the defective transposition functions of Mud1-8.