Increased stability of nucleic acids containing 7-deaza-guanosine and 7-deaza-adenosine may enable rapid DNA sequencing by matrix-assisted laser desorption mass spectrometry.
Increased stability of nucleic acids containing 7-deaza-guanosine and 7-deaza-adenosine may enable rapid DNA sequencing by matrix-assisted laser desorption mass spectrometry.
复制标题
含有 7-脱氮鸟苷和 7-脱氮腺苷的核酸的稳定性增加可以通过基质辅助激光解吸质谱法实现快速 DNA 测序。
DOI：
10.1093/nar/23.9.1570
复制
发表时间：
1995
影响因子：
14.9
通讯作者：
Chait,BT
中科院分区：
文献类型：
--
作者：
Schneider,K;Chait,BT
基质辅助激光解吸质谱仪(MALDI-MS)的使用被认为是Sanger DNA测序梯子的超快读数，其方式类似于测序凝胶使用的方式。目前，MALDI-MS用于DNA分析的严重局限性是由于寡核苷酸在气相中容易发生碎裂的趋势造成的。本研究旨在了解由MALDI产生的寡聚脱氧核苷酸离子的各种化学结构特征对其稳定性的影响。研究了d型(TTTTTTTTTTXTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT核苷X中选择了多种不同的嘌呤类化合物作为碱基。在MALDI-MS的条件下，含有7-去氮鸟嘌呤和腺苷类似物的模型化合物的质谱图显示出比7-氮杂类似物显著增加的稳定性。先前报道的7-脱氮-2‘-脱氧腺苷三磷酸和7-脱氮-2’-脱氧鸟苷三磷酸通过聚合酶结合到DNA中，这表明它们可用于Sanger双脱氧测序实验。用7-脱氮嘌呤代替7-氮杂嘌呤的双脱氧终止产物可能足够稳定，从而可以用MALDI-MS分离和检测测序阶梯。因此，DNA序列的超快(秒)读出可能成为可能。
The use of matrix-assisted laser desorption mass spectrometry (MALDI-MS) has been suggested as an ultrafast readout of Sanger DNA sequencing ladders in a manner analogous to that used with sequencing gels. Currently, a serious limitation of MALDI-MS for the analysis of DNA results from the tendency for oligonucleotides to undergo facile fragmentation in the gas phase. The present study was undertaken to gain an understanding of the influence of various chemical structural features of purlne bases on the stability of oligodeoxynucleotide ions produced by MALDI. The study focused on the stability of model compounds of the type d(TTTTTTTTTTXTTTTTTTTTTTTT), where T designates deoxythymldlne and X a purine-contalning 2′-deoxynucleotide. A variety of different purine derivatives were chosen as the base in the nucleotlde X. The mass spectra of the model compounds containing 7-deaza analogues of guanine and adenlne reveal a significantly increased stability compared to the 7-aza analogues under the conditions of MALDI-MS. The previously reported incorporation of the 7-deaza-2′-deoxy-adenosine trlphosphate and the 7-deaza-2′-deoxy-guanosine triphosphate into DNA by polymerases suggests their use in a Sanger dideoxy sequencing experiment. The dideoxy termination products with the 7-deaza-purines instead of the 7-aza-purines might be sufficiently stable to allow separation and detection of the sequencing ladder by MALDI-MS. Thus, an ultrafast (seconds) read-out of DNA sequence may become feasible.