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淡紫灰链霉菌66基因组的物理图谱及其与相关菌株天蓝色链霉菌A3(2)基因组的比较。

Physical map of the Streptomyces lividans 66 genome and comparison with that of the related strain Streptomyces coelicolor A3(2).

作者信息

Leblond P, Redenbach M, Cullum J

机构信息

LB Genetik der Universität Kaiserslautern, Germany.

出版信息

J Bacteriol. 1993 Jun;175(11):3422-9. doi: 10.1128/jb.175.11.3422-3429.1993.

Abstract

A physical map of the chromosome of Streptomyces lividans 66 ZX7 was constructed by ordering the macrorestriction fragments generated from the genomic DNA with the restriction enzymes AseI and DraI. AseI and DraI linking cosmids (i.e., recombinant cosmids including either AseI or DraI sites) were isolated from a gene bank and used as hybridization probes against Southern transfers of pulsed-field gel electrophoresis (PFGE) restriction patterns. The DraI sites were precisely mapped by PFGE analyses of AseI-DraI double digests and hybridization with the AseI junctions. The 16 AseI and 7 DraI fragments were aligned as a single chromosome of about 8,000 kb. The data supported the interpretation that the chromosome is a linear structure. The related strain Streptomyces coelicolor A3(2) M145, recently mapped by H. Kieser, T. Kieser, and D. A. Hopwood (J. Bacteriol. 174:5496-5507, 1992), was compared with S. lividans at the level of the genomic structure by hybridizing the linking cosmids to Southern transfers of PFGE patterns. In spite of little apparent similarity in their restriction patterns, the comparison of the physical maps revealed a common structure with an identical ordering of the cosmid sequences. This conservation of the map order was further confirmed by assigning genetic markers (i.e., cloned genes and DNA elements relevant to the unstable region) to the AseI fragments.

摘要

通过对用限制性内切酶AseI和DraI从基因组DNA产生的大限制性片段进行排序,构建了变铅青链霉菌66 ZX7染色体的物理图谱。从基因文库中分离出连接黏粒的AseI和DraI(即包含AseI或DraI位点的重组黏粒),并将其用作针对脉冲场凝胶电泳(PFGE)限制性图谱的Southern转移杂交探针。通过对AseI-DraI双酶切产物进行PFGE分析并与AseI接头杂交,精确绘制了DraI位点。16个AseI片段和7个DraI片段排列成一条约8000 kb的单染色体。这些数据支持了染色体是线性结构的解释。通过将连接黏粒与PFGE图谱的Southern转移杂交,在基因组结构水平上比较了相关菌株天蓝色链霉菌A3(2) M145(最近由H. Kieser、T. Kieser和D. A. Hopwood绘制图谱,《细菌学杂志》174:5496 - 5507,1992年)和变铅青链霉菌。尽管它们的限制性图谱几乎没有明显相似性,但物理图谱的比较揭示了一种共同结构,其中黏粒序列的排序相同。通过将遗传标记(即克隆基因和与不稳定区域相关的DNA元件)定位到AseI片段,进一步证实了图谱顺序的保守性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9295/204741/6c1417d863fe/jbacter00053-0190-a.jpg

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