Patch Ann-Marie, Aves Stephen J
School of Biosciences, University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
Microbiology (Reading). 2007 Mar;153(Pt 3):887-97. doi: 10.1099/mic.0.2006/001669-0.
The fission yeast Schizosaccharomyces pombe is widely used as a model eukaryote for cell and molecular studies but little is known of natural genetic variation in this species. In order to obtain informative molecular markers, imperfect tandem repeats, identified through bioinformatic methods, were tested for length polymorphism in six wild-type strains of Sch. pombe isolated from different substrates and geographical locations in Africa, America, Asia and Europe. Of 26 loci tested, 21 were multi-allelic, consistent with tandem repeat copy number variation. Eleven of these polymorphic tandem repeats are in regions encoding intracellular proteins. Most of the protein-coding repeats are not sited within structured domains but have non-regular predicted structure; one has a repeat unit length corresponding to integer turns of a predicted amphipathic alpha-helix secondary structure, suggesting that this repeat may be tolerated because copy number mutations change alpha-helix length but not orientation within the protein structure. In contrast to the differences observed between natural isolates of Sch. pombe, genetic strains were found to be essentially isogenic: only two polymorphic loci were detected out of 26 minisatellites and five microsatellites tested in 16 strains, including a hypervariable microsatellite in the med15 gene. The polymorphic tandem repeat markers identified in this study will prove useful for DNA fingerprinting and molecular analysis of natural genetic variation in Sch. pombe isolates.
裂殖酵母粟酒裂殖酵母被广泛用作细胞和分子研究的真核生物模型,但对该物种的自然遗传变异了解甚少。为了获得有用的分子标记,通过生物信息学方法鉴定出的不完全串联重复序列,在从非洲、美洲、亚洲和欧洲不同底物和地理位置分离的6株粟酒裂殖酵母野生型菌株中进行了长度多态性测试。在测试的26个位点中,21个是多等位基因的,与串联重复拷贝数变异一致。其中11个多态性串联重复序列位于编码细胞内蛋白质的区域。大多数蛋白质编码重复序列不在结构化结构域内,但具有不规则的预测结构;其中一个重复单元长度对应于预测的两亲性α-螺旋二级结构的整数圈,这表明该重复序列可能被容忍,因为拷贝数突变会改变α-螺旋长度,但不会改变蛋白质结构内的方向。与粟酒裂殖酵母自然分离株之间观察到的差异相反,发现遗传菌株基本同基因:在测试的16株菌株中的26个小卫星和5个微卫星中,仅检测到2个多态性位点,包括med15基因中的一个高变微卫星。本研究中鉴定的多态性串联重复标记将被证明可用于粟酒裂殖酵母分离株的DNA指纹识别和自然遗传变异的分子分析。