Zhao Y, Zhang S
Department of Medical Genetics, The Affiliated Hospital, West China University of Medical Sciences, Chengdu, Sichuan, 610041 P.R.China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 1999 Oct;16(5):333-6.
To introduce a new method for studies on methylation status of CpG island: methylation specific polymerase chain reaction (MSP) with some modifications.
Target DNA was modified by sodium bisulfite, converting all unmethylated, but not methylated, cytodines to uracil, and subsequent amplification with primers specific for methylated versus unmethylated DNA. The status of 5'CpG island of tumor suppressor gene p16 in maxilliofacial squamous cell carcinoma (MSCC) was analyzed by modified MSP assay.
To get single strand DNA, the authors denatured the target DNA by heating instead of alkaline treatment. DNA purification kit was also replaced by dialysis. A cheaper restriction enzyme Taq I was used in place of BstU I to distinguish methylated fragment from unmethylated fragment. The 5'CpG island of p16 gene was methylated in some MSCC tumors.
MSP is a simple, sensitive, and specific method for detecting the methylation status of any CpG-rich region and the modifications make it more simple to perform.
介绍一种研究CpG岛甲基化状态的新方法：改良的甲基化特异性聚合酶链反应（MSP）。
用亚硫酸氢钠修饰靶DNA，将所有未甲基化的胞嘧啶（而非甲基化的胞嘧啶）转化为尿嘧啶，随后用针对甲基化与未甲基化DNA的引物进行扩增。通过改良的MSP分析法分析口腔颌面部鳞状细胞癌（MSCC）中抑癌基因p16的5'CpG岛状态。
为获得单链DNA，作者通过加热而非碱处理使靶DNA变性。还用透析代替了DNA纯化试剂盒。使用更便宜的限制性内切酶Taq I代替BstU I来区分甲基化片段和未甲基化片段。p16基因的5'CpG岛在一些MSCC肿瘤中发生了甲基化。
MSP是一种检测任何富含CpG区域甲基化状态的简单、灵敏且特异的方法，这些改良使其操作更加简便。
