Cloning of Lentinus edodes mitochondrial DNA fragment capable of autonomous replication in Saccharomyces cerevisiae.
Cloning of Lentinus edodes mitochondrial DNA fragment capable of autonomous replication in Saccharomyces cerevisiae.
复制标题
能够在酿酒酵母中自主复制的香菇线粒体DNA片段的克隆。
DOI：
10.1016/s0006-291x(86)80397-7
复制
发表时间：
1986
期刊：
影响因子：
--
通讯作者：
M. Ohmasa
中科院分区：
文献类型：
--
作者：
Y. Katayose;K. Shishido;M. Ohmasa
用sau3ai酶切了分子量约为69 kb的高等承载菌lentinus edodeses的线粒体（mt） DNA，用质粒YIp32（pBR322与酵母基因yeastle2的杂交产物）进行克隆，并在真核糖酵母酿酒酵母（cerevisiae）中分析了具有自主复制（ARSs）能力的序列。重组质粒中含有j . 2kb具有ARS活性的mtDNA片段。该质粒（命名为pSK52）表现出高频酵母转化，并被发现作为染色体外元件维持在细胞内。pSK52的稳定性和拷贝数特性与YIp32的重组质粒相似。cerevisiaem -作为引用构造的ars。通过亚克隆实验对上述3.2 kb片段的ARS定位进行了评估，发现该片段至少包含两个ARS。
Mitochondrial(mt) DNA of the higher basidiomycetesLentinus edodeswith a molecular weight of about 69 kb was partially digested withSau3AI, cloned with plasmid YIp32(a hybrid of pBR322 and the yeastleu2 gene) and analyzed for sequences capable of autonomous replication(ARSs) in the eukaryoteSaccharomyces cerevisiae. One recombinant plasmid was isolated which contained J.2 kb fragment of the mtDNA with ARS activity. This plasmid(named pSK52) exhibited a high-frequency yeast transformation and was found to be maintained within the cell as an extrachromosomal element. The stability and copy number properties of pSK52 were similar to those of the recombinant plasmid of YIp32 andS. cerevisiaemt-ARS constructed as a reference. Subcloning experiments were carried out to assess the localization of ARS on the above 3.2 kb fragment, revealing that the fragment contains at least two ARSs.