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嗜热栖热菌HB8质粒克隆载体的构建:异源质粒携带的卡那霉素核苷酸转移酶基因的表达

Development of plasmid cloning vectors for Thermus thermophilus HB8: expression of a heterologous, plasmid-borne kanamycin nucleotidyltransferase gene.

作者信息

Mather M W, Fee J A

机构信息

Biochemistry Section, Los Alamos National Laboratory, New Mexico 87545.

出版信息

Appl Environ Microbiol. 1992 Jan;58(1):421-5. doi: 10.1128/aem.58.1.421-425.1992.

Abstract

While several Thermus genes have been cloned and T. thermophilus has been shown to be transformable, molecular genetic studies of these thermophiles have been hampered by the absence of selectable cloning vectors. We have constructed a selectable plasmid by random insertion of a heterologous gene encoding a thermostable kanamycin nucleotidyltransferase activity into a cryptic, multicopy plasmid from T. thermophilus HB8. This plasmid should serve as a suitable starting point for the development of a gene expression system for T. thermophilus.

摘要

虽然已经克隆了几个嗜热栖热菌基因,并且嗜热栖热菌已被证明具有可转化性,但由于缺乏可选择的克隆载体,这些嗜热菌的分子遗传学研究受到了阻碍。我们通过将编码热稳定卡那霉素核苷酸转移酶活性的异源基因随机插入来自嗜热栖热菌HB8的一个隐蔽多拷贝质粒中,构建了一个可选择的质粒。该质粒应可作为开发嗜热栖热菌基因表达系统的合适起点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8321/195228/1e4af545b136/aem00042-0443-a.jpg

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