Cheung Iris, Schertzer Mike, Baross Agnes, Rose Ann M, Lansdorp Peter M, Baird Duncan M
Terry Fox Laboratory, BC Cancer Agency, Vancouver, BC V5Z 4E6, Canada.
Nucleic Acids Res. 2004 Jun 24;32(11):3383-91. doi: 10.1093/nar/gkh661. Print 2004.
Terminal restriction fragment analysis is the only method currently available for measuring telomere length in Caenorhabditis elegans. Its limitations include low sensitivity and interference by the presence of interstitial telomeric sequences in the C.elegans genome. Here we report the adaptation of single telomere length analysis (STELA) to measure the length of telomeric repeats on the left arm of chromosome V in C.elegans. This highly sensitive PCR-based method allows telomere length measurement from as few as a single worm. The application of STELA to eight wild-type C.elegans strains revealed considerable strain-specific differences in telomere length. Within individual strains, short outlying telomeres were observed that were clearly distinct from the bulk telomere length distributions, suggesting that processes other than end-replication losses and telomerase-mediated lengthening may generate telomere length heterogeneity in C.elegans. The utility of this method was further demonstrated by the characterization of telomere shortening in mrt-2 mutants. We conclude that STELA appears to be a valuable tool for studying telomere biology in C.elegans.
末端限制片段分析是目前唯一可用于测量秀丽隐杆线虫端粒长度的方法。其局限性包括灵敏度低以及秀丽隐杆线虫基因组中存在的间质端粒序列的干扰。在此,我们报告了单端粒长度分析(STELA)的改进方法,用于测量秀丽隐杆线虫第五条染色体左臂上的端粒重复序列长度。这种基于PCR的高灵敏度方法能够从仅一条线虫中测量端粒长度。将STELA应用于八个野生型秀丽隐杆线虫菌株,结果显示端粒长度存在显著的菌株特异性差异。在各个菌株中,观察到了短的异常端粒,它们明显不同于大部分端粒长度分布,这表明除了末端复制损耗和端粒酶介导的延长之外,其他过程可能导致秀丽隐杆线虫中端粒长度的异质性。通过对mrt-2突变体中端粒缩短的表征,进一步证明了该方法的实用性。我们得出结论,STELA似乎是研究秀丽隐杆线虫端粒生物学的一个有价值的工具。