Haplotype mapping of a diploid non-meiotic organism using existing and induced aneuploidies.
Haplotype mapping of a diploid non-meiotic organism using existing and induced aneuploidies.
复制标题
使用现有的和诱导的非整倍性化的二倍体非舒张生物的单倍型映射。
DOI：
10.1371/journal.pgen.0040001
复制
发表时间：
2008-01
期刊：
影响因子：
4.5
通讯作者：
Berman, Judith
中科院分区：
文献类型：
--
作者：
Legrand, Melanie;Forche, Anja;Selmecki, Anna;Chan, Christine;Kirkpatrick, David T.;Berman, Judith
单倍型图谱（HapMap）揭示了潜在的序列变异，并促进重组和遗传多样性的研究。一般来说，单体型图是通过分析大量减数分裂后代中的单核苷酸多态性 (SNP) 分离而产生的。白色念珠菌是最常见的人类真菌病原体，是一种专性二倍体，似乎不会经历减数分裂。因此，不能使用单倍型映射的标准方法。我们利用天然存在的非整倍体菌株来确定白色念珠菌实验室菌株 SC5314 和临床分离株中八个染色体对的单倍型。图谱的比较表明，临床菌株经历了大量的基因组重排，主要包括交叉或基因转换重组事件。 SNP 图谱单倍型分析显示，必需和非必需基因中 UAU1 盒的插入和激活可导致整个染色体非整倍性。 UAU1 通常用于构建白色念珠菌中目标基因的纯合缺失；确切的机制（三体性随后染色体丢失与基因转换）尚未确定。 UAU1 插入必需的 ORC1 基因会导致大部分三体菌株，而当 UAU1 插入非必需的 LRO1 基因时，基因转换事件占主导地位。因此，诱导的非整倍体可用于生成 HapMap，这对于分析该克隆生物体中的基因组改变和有丝分裂重组事件至关重要。 白色念珠菌是一种杂合二倍体酵母，是最常见的真菌病原体。它通常通过基因组改变重组事件获得抗真菌药物的耐药性。在许多生物体中，重组事件是使用单倍型图谱 (HapMaps) 进行分析的，它显示了每个染色体同源物上不同等位基因的位置。传统的单体型图是通过在减数分裂后代中分离时遵循等位基因标记来构建的。由于白色念珠菌尚未被证明会经历减数分裂，因此不可能构建念珠菌单体型图。我们利用白色念珠菌有丝分裂后代中全染色体非整倍体的存在来检测不同等位基因的倾斜比率，从而确定每个染色体同源物上这些等位基因之间的关系。这有助于构建最常用的白色念珠菌实验室菌株的 HapMap。然后，我们使用该 HapMap 来识别临床分离株中相对于实验室参考菌株的所有重组事件。最后，我们使用这种作图方法来研究当白色念珠菌基因组受到常见基因破坏技术时影响其基因组的分子机制。我们的快速单体型图构建方法通常适用于可以识别全染色体非整倍性事件的任何生物体。
Haplotype maps (HapMaps) reveal underlying sequence variation and facilitate the study of recombination and genetic diversity. In general, HapMaps are produced by analysis of Single-Nucleotide Polymorphism (SNP) segregation in large numbers of meiotic progeny. Candida albicans, the most common human fungal pathogen, is an obligate diploid that does not appear to undergo meiosis. Thus, standard methods for haplotype mapping cannot be used. We exploited naturally occurring aneuploid strains to determine the haplotypes of the eight chromosome pairs in the C. albicans laboratory strain SC5314 and in a clinical isolate. Comparison of the maps revealed that the clinical strain had undergone a significant amount of genome rearrangement, consisting primarily of crossover or gene conversion recombination events. SNP map haplotyping revealed that insertion and activation of the UAU1 cassette in essential and non-essential genes can result in whole chromosome aneuploidy. UAU1 is often used to construct homozygous deletions of targeted genes in C. albicans; the exact mechanism (trisomy followed by chromosome loss versus gene conversion) has not been determined. UAU1 insertion into the essential ORC1 gene resulted in a large proportion of trisomic strains, while gene conversion events predominated when UAU1 was inserted into the non-essential LRO1 gene. Therefore, induced aneuploidies can be used to generate HapMaps, which are essential for analyzing genome alterations and mitotic recombination events in this clonal organism. Candida albicans, a heterozygous diploid yeast, is the most prevalent fungal pathogen. It often acquires resistance to anti-fungal drugs via genome-altering recombination events. In many organisms, recombination events are analyzed using Haplotype Maps (HapMaps), which show the location of different alleles on each chromosomal homolog. Conventional HapMaps are constructed by following allelic markers as they segregate in meiotic progeny. Because C. albicans has not been shown to undergo meiosis, construction of a Candida HapMap has not been possible. We exploited the presence of whole chromosome aneuploidies in mitotic progeny of C. albicans to detect skewed ratios of different alleles, thereby determining the relationships between these alleles on each chromosomal homolog. This facilitated the construction of a HapMap for the most commonly used C. albicans laboratory strain. We then used this HapMap to identify all of the recombination events in a clinical isolate relative to the laboratory reference strain. Finally, we used this mapping approach to investigate the molecular mechanisms that affect the C. albicans genome when it is subjected to a common gene disruption technique. Our rapid HapMap construction method is generally applicable to any organism for which whole-chromosome aneuploidy events can be identified.
登录
查看更多内容
影响因子：
3.2
作者：
Cowen, LE;Sanglard, D;Kohn, LM
通讯作者：
Kohn, LM
影响因子：
2.8
作者：
Wellington, Melanie;Kabir, M. Anaul;Rustchenko, Elena
通讯作者：
Rustchenko, Elena
影响因子：
3.6
作者：
Legrand, M;Lephart, P;Magee, BB
通讯作者：
Magee, BB
影响因子：
64.8
作者：
Gibbs, RA;Belmont, JW;Tanaka, T
通讯作者：
Tanaka, T
影响因子：
3.2
作者：
Wilson, RB;Davis, D;Mitchell, AP
通讯作者：
Mitchell, AP