巨大线性质粒SCP1的EcoRI片段的克隆与物理图谱构建
Cloning and physical mapping of the EcoRI fragments of the giant linear plasmid SCP1.
作者信息
Redenbach M, Ikeda K, Yamasaki M, Kinashi H
机构信息
Department of Molecular Biotechnology, Graduate School of Engineering, Hiroshima University, Higashi-Hiroshima 739-8527, Japan.
出版信息
J Bacteriol. 1998 May;180(10):2796-9. doi: 10.1128/JB.180.10.2796-2799.1998.
Abstract
A cosmid library was constructed for the 350-kb giant linear plasmid SCP1 and aligned on a successive linear map. Only a 0.8-kb gap has remained uncloned in the terminal inverted repeats close to both ends. Partial digestion of the aligned cosmids with EcoRI and hybridization with the flanking fragments of the vector enabled physical mapping of all of the EcoRI fragments. On this map, the methylenomycin biosynthetic gene cluster, the insertion sequence IS466, and the sapCDE genes coding for spore-associated proteins were localized.
摘要
构建了一个针对350kb巨型线性质粒SCP1的黏粒文库，并将其排列在连续的线性图谱上。在靠近两端的末端反向重复序列中，仅剩下一个0.8kb的缺口未被克隆。用EcoRI对排列好的黏粒进行部分酶切，并与载体的侧翼片段杂交，从而实现了所有EcoRI片段的物理图谱绘制。在该图谱上，定位了亚甲基霉素生物合成基因簇、插入序列IS466以及编码孢子相关蛋白的sapCDE基因。
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