Egilmez N K, Ebert R H, Shmookler Reis R J
Department of Medicine, University of Arkansas for Medical Sciences, Little Rock, USA.
J Mol Evol. 1995 Apr;40(4):372-81. doi: 10.1007/BF00164023.
Evolutionary relationships across taxa can be deduced from sequence divergence of proteins, RNA, or DNA; sequences which diverge rapidly, such as those of mitochondrial genes, have been especially useful for comparisons of closely related species, and--within limits--of strains within a species. We have utilized the transposable element Tc1 as a polymorphic marker to evaluate the evolutionary relationships among nine Caenorhabditis elegans strains. For five low-Tc1-copy strains, we compared patterns of restriction fragments hybridizing to a cloned Tc1 probe. Twenty of the 40 Tc1 insertion sites thus characterized were common to all five strains, and so presumably preceded strain divergence; the 20 differential bands were used to construct a maximum-parsimony tree relating these strains. In four high-copy-number stocks (three wild-type strains and a subline), we determined occupancy of 35 individual Tc1 insertion sites by a polymerase chain reaction assay. Surprisingly, the high-copy strains share a common subset of these Tc1 insertions, and the chromosomal distribution of conserved Tc1 sites is "clustered" with respect to the other elements tested. These data imply a close evolutionary relationship among the high-copy strains, such that two of these strains appear to have been derived from the highest-copy-number lineage (represented by two stocks) through crossing with a low-Tc1 strain. Abundances of Tc1 elements were also estimated for the four high-copy-number stocks, at approximately 200-500 copies per haploid genome, by quantitative dot-blot hybridization relative to two low-copy strains. Annealing with 32P-labeled probes corresponding to full-length Tc1, an oligonucleotide within the Tc1 terminal inverted repeats, and an internal Tc1 oligonucleotide, gave essentially identical results--indicating that Tc1 termini exist in the genome primarily as components of full-length Tc1 elements. A composite evolutionary tree is proposed, based on the locations and numbers of Tc1 elements in these strains, which is consistent with a four-branch intraspecific tree deduced previously by maximum-parsimony analyses of mitochondrial sequence changes; it also serves to elucidate the evolutionary history of transposon mobility.
不同分类单元之间的进化关系可以从蛋白质、RNA或DNA的序列差异中推导出来;快速分化的序列,如线粒体基因序列,对于比较亲缘关系密切的物种,以及在一定限度内比较同一物种内的菌株特别有用。我们利用转座因子Tc1作为多态性标记来评估9种秀丽隐杆线虫菌株之间的进化关系。对于5个低Tc1拷贝数的菌株,我们比较了与克隆的Tc1探针杂交的限制性片段模式。在如此鉴定出的40个Tc1插入位点中,有20个为所有5个菌株所共有,因此推测在菌株分化之前就已存在;这20条差异带被用于构建一棵最大简约树,以关联这些菌株。在4个高拷贝数品系(3个野生型菌株和1个亚系)中,我们通过聚合酶链反应分析确定了35个单个Tc1插入位点的占据情况。令人惊讶的是,高拷贝数菌株共享这些Tc1插入的一个共同子集,并且保守的Tc1位点的染色体分布相对于所测试的其他元件是“聚集的”。这些数据表明高拷贝数菌株之间存在密切的进化关系,以至于其中两个菌株似乎是通过与一个低Tc1菌株杂交,从最高拷贝数谱系(由两个品系代表)衍生而来。通过相对于两个低拷贝菌株的定量点杂交,还估计了4个高拷贝数品系中Tc1元件的丰度,大约为每个单倍体基因组200 - 500个拷贝。用对应于全长Tc1的32P标记探针、Tc1末端反向重复序列内的一个寡核苷酸以及一个内部Tc1寡核苷酸进行退火,得到了基本相同的结果——表明Tc1末端在基因组中主要作为全长Tc1元件的组成部分存在。基于这些菌株中Tc1元件的位置和数量,提出了一棵复合进化树,它与先前通过线粒体序列变化的最大简约分析推导的一棵四分支种内树一致;它也有助于阐明转座子移动性的进化历史。