Accurate characterization of the tyrosine hydroxylase forensic allele 9.3 through development of electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry.
Accurate characterization of the tyrosine hydroxylase forensic allele 9.3 through development of electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry.
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通过开发电喷雾电离傅里叶变换离子回旋共振质谱法准确表征酪氨酸羟化酶法医等位基因 9.3。
DOI：
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发表时间：
1999
影响因子：
2
通讯作者：
D. Muddiman
中科院分区：
文献类型：
--
作者：
J. Hannis;D. Muddiman
首次通过电喷雾电离傅里叶变换离子回旋共振质谱（ESI-FTICR-MS），准确和精确地测定了人类基因位点短串联重复序列（STR）的重复数。具体而言，多态性的人酪氨酸羟化酶（HUMTHO 1）基因，四核苷酸STR法医等位基因，被选为一个模型系统，以评估我们的方法，为未来表征STR和可变数目的串联重复序列（VNTRs）的ESI-FTICR-MS方法的发展。同时解析来自HUMTHO 1 9.3等位基因的编码链和非编码链，获得编码链和非编码链分别为25，783.23和24，754.55 Da的准确（优于70 ppm）平均质量测量值。质量测量用于计算每条链的重复数，“n”，编码链和非编码链分别为9.75169和9.75001。它将显示“n”的值如何可以用来直接确定纯重复的数量，并准确地确定重复内的多态性的确切性质（如果有的话）。使用这种方法准确地鉴定了编码链（腺嘌呤）和非编码链（胸腺嘧啶）中的单核苷酸缺失。有趣的是，我们观察到单链DNA转化为双链DNA，而在ESI缓冲液中的PCR产物被注入;与此相关的问题将被提出。其他研究人员使用基质辅助激光解吸/电离飞行时间（MALDI-TOF）研究HUMTHO 1 9.3等位基因的先前结果与我们的结果直接比较。我们的研究结果表明，ESI-FTICR-MS是一种强大的方法，快速，准确地表征串联重复序列，这将最终导致对一类复杂的疾病的理解和人类身份的确定。
Accurate and precise determination of the number of repeats from a short tandem repeat (STR) sequence for a human gene locus is demonstrated for the first time by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR-MS). Specifically, the polymorphic human tyrosine hydroxylase (HUMTHO1) gene, a tetranucleotide STR forensic allele, was chosen as a model system to evaluate our approach for future characterization of both STRs and variable number of tandem repeats (VNTRs) by development of an ESI-FTICR-MS approach. The coding and noncoding strands from the HUMTHO1 9.3 allele are simultaneously resolved obtaining accurate (better than 70 ppm) average mass measurements of 25,783.23 and 24,754.55 Da for the coding and noncoding strands, respectively. The mass measurements are used to calculate the number of repeats for each strand, 'n', of 9.75169 and 9.75001 for the coding and noncoding strands, respectively. It will be shown how the value of 'n' can be used to directly determine the number of pure repeats and accurately determine the exact nature of the polymorphism within the repeat (if any). The single nucleotide deletion in the coding strand (adenine) and noncoding strand (thymine) were accurately identified using this approach. Interestingly, we observed the conversion of single-stranded to double-stranded DNA while the PCR product in the ESI buffer was being infused; the issues related to this observation will be presented. Previous results by other researchers investigating the HUMTHO1 9.3 allele using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) are directly compared with our results. Our results indicate that ESI-FTICR-MS is a powerful approach to rapidly and accurately characterize tandem repeating sequences which will ultimately lead towards the understanding of a complex class of diseases and in human identity determination.
影响因子：
4.4
作者：
EDWARDS, A;HAMMOND, HA;CHAKRABORTY, R
通讯作者：
CHAKRABORTY, R
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。
影响因子：
14.9
作者：
Schneider,K;Chait,BT
通讯作者：
Chait,BT