基于质谱的核酸指纹识别技术

Nucleic acid fingerprint technique for RNA analysis based mass spectrometry

基因转录是继基因组计划完成后基因功能研究的基本内容。基因转录研究迫切需要有新方法的支持。人们从蛋白的指纹分析技术中得到启发,开发基于质谱分析的核酸指纹识别技术。通过对核酸进行酶解处理,使其核酸片段分子量位于质谱检测的范围内,通过高分辨质谱技术获得所检测酶解核酸片段的分子量列表;利用基因组研究所获得的大量数据,通过计算机技术,模拟产生核酸酶解产物数据库,通过独特的算法以及基于这些算法开发的工具软件,把酶解的核酸质谱数据和虚拟的核酸酶解产物数据库进行比对,以发现和鉴别核酸。随着这一技术的逐渐应用和发展,将来还可能直接建立基于质谱数据的核酸数据库。相信这一技术在转录组研究中将发挥重要作用。

Genetic transcription, as the first stage of gene function study, is the hotpot of post-genomic research. Genetic transcription study required support from development of brand-new methods. Recently mass spectrometry based nucleic acid fingerprint technique was successfully developed for RNA analysis, which learned from the peptide mass finger technique widely used in protein analysis. High molecular weight RNA was cut by nuclease to low molecular weight fragments for high resolution mass spectrometry （MS） analysis, then the acquired mass list from mass spectrometry was loaded to software for evaluating the matches between the masses of product ions generated by MS and in silico database calculated from a candidate nucleotide sequence in a DNA/RNA sequence database, predicting the nucleotide sequences of those RNase fragments. Joining those fragments together, scoring or ranking of all matches, the final RNA would be identified and mapped to genome. Mass spectrometry based RNA database as substitution of above in silico database can be used with further development of this technique. I believe this strategy will be used widely in genetic transcription area.