Efficient homologous and illegitimate recombination in the opportunistic yeast pathogen Candida glabrata.
Efficient homologous and illegitimate recombination in the opportunistic yeast pathogen Candida glabrata.
复制标题
机会性酵母病原体光滑念珠菌中的高效同源和非法重组。
作者：
B. Cormack;S. Falkow;S. Falkow
条件致病菌光滑假丝酵母菌在人类中会引起重大疾病。为了开发基因工具来研究这种微生物的致病性，我们通过删除光滑毛囊线虫临床分离株的相关编码区，构建了ura3和his3营养缺陷型菌株。携带酿酒酵母URA3基因的线性化载体能有效地将ura3营养缺陷菌转化为原生生物。当线性化的质粒携带与染色体同源的短末端区域时，观察到同源重组事件。相反，在没有任何染色体同源性的情况下，通过不合法的重组将质粒整合到基因组中的随机位置。对目标位点的序列分析显示，对于大多数非法转化子，与整合位点没有微同源性。大约0.25%的插入导致氨基酸营养不良，这表明插入在总体水平上是随机的。序列分析表明，非法重组在单基因水平上是非随机的，整合的质粒倾向于插入基因组的非编码区。对同源和非同源重组事件的相对数量的分析表明，光滑毛壳菌具有有效的同源和非同源重组系统。
The opportunistic pathogen Candida glabrata causes significant disease in humans. To develop genetic tools to investigate the pathogenicity of this organism, we have constructed ura3 and his3 auxotrophic strains by deleting the relevant coding regions in a C. glabrata clinical isolate. Linearized plasmids carrying a Saccharomyces cerevisiae URA3 gene efficiently transformed the ura3 auxotroph to prototrophy. Homologous recombination events were observed when the linearized plasmid carried short terminal regions homologous with the chromosome. In contrast, in the absence of any chromosomal homology, the plasmid integrated by illegitimate recombination into random sites in the genome. Sequence analysis of the target sites revealed that for the majority of illegitimate transformants there was no microhomology with the integration site. Approximately 0.25% of the insertions resulted in amino acid auxotrophy, suggesting that insertion was random at a gross level. Sequence analysis suggested that illegitimate recombination is nonrandom at the single-gene level and that the integrating plasmid has a preference for inserting into noncoding regions of the genome. Analysis of the relative numbers of homologous and illegitimate recombination events suggests that C. glabrata possesses efficient systems for both homologous and nonhomologous recombination.
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使用来自金属硫蛋白-IIa 编码基因的元件的光滑念珠菌克隆系统，该基因赋予自主复制能力。
影响因子：
3.5
作者：
Mehra,RK;Thorvaldsen,JL;Macreadie,IG;Winge,DR
通讯作者：
Winge,DR
用于酵母光滑念珠菌基因克隆和操作的系统。
影响因子：
3.5
作者：
Zhou,P;Szczypka,MS;Young,R;Thiele,DJ
通讯作者：
Thiele,DJ
影响因子：
14.9
作者：
BEEN, MD;BURGESS, RR;CHAMPOUX, JJ
通讯作者：
CHAMPOUX, JJ
DOI：
10.1073/pnas.88.17.7585
发表时间：
1991-09-01
影响因子：
11.1
作者：
SCHIESTL, RH;PETES, TD
通讯作者：
PETES, TD