CLONING OF 18S AND 25S RDNAS FROM THE PATHOGENIC FUNGUS CRYPTOCOCCUS-NEOFORMANS
CLONING OF 18S AND 25S RDNAS FROM THE PATHOGENIC FUNGUS CRYPTOCOCCUS-NEOFORMANS
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DOI：
10.1128/jb.171.10.5596-5600.1989
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发表时间：
1989-10-01
影响因子：
3.2
通讯作者：
BARBOUR, AG
中科院分区：
文献类型：
--
作者：
RESTREPO, BI;BARBOUR, AG
新型隐球菌是一种重要的病原真菌，已被归类为担子菌。这种真菌病原体的分子遗传学知之甚少。为了开始这样的研究，我们设计了一种从隐球菌中提取DNA的方法;这种方法涉及使用细胞壁活性酶NovoZym 234。以克隆的酿酒酵母rDNA为探针，对C.新生儿DNA将与酵母rDNA探针杂交的8.6-内切酶HindIII片段与载体pBR 322连接并克隆到大肠杆菌中。以该片段为探针，与C.在天然和变性RNA的北方（RNA）印迹中，它在高严格性下仅与子囊菌S的rRNA弱结合。啤酒。用这些不同类型的标记rRNA鉴定了克隆片段中5/5.8S、18 S和25 S亚基基因的位置。
Cryptococcus neoformans is an important pathogenic fungus that has been classified as a basidiomycete. Little is known of the molecular genetics of this fungal pathogen. To begin such studies, we devised a procedure for extraction of DNA from cryptococci; this method involved the use of the cell wall-active enzyme NovoZym 234. Using cloned rDNA of Saccharomyces cerevisiae as a probe, we identified homologous restriction fragments in a Southern blot of digested C. neoformans DNA. An 8.6-kilobase HindIII fragment that hybridized with the yeast rDNA probe was ligated with the vector pBR322 and cloned into Escherichia coli. When the fragment was used as a probe, it hybridized to the 18S and 25S rRNAs of C. neoformans in Northern (RNA) blots of native and denatured RNA. It bound at high stringency only weakly to the rRNAs of the ascomycete S. cerevisiae. The locations of the genes for 5/5.8S, 18S, and 25S subunits in the cloned fragment were identified with labeled rRNA of these different types.