详细信息
三苯甲基保护的巯烷臂寡核苷酸的合成及荧光标记
Synthesis of an Oligonucleotide Probe Containing Trityl-protected Sulphydryl Group and Tagging with a Fluorescent Dye
文献类型:期刊文献
中文题名:三苯甲基保护的巯烷臂寡核苷酸的合成及荧光标记
英文题名:Synthesis of an Oligonucleotide Probe Containing Trityl-protected Sulphydryl Group and Tagging with a Fluorescent Dye
作者:魏欣[1];张惠展[1];黎占亭[2];姚伟[3];钱旭红[3]
机构:[1]华东理工大学生物反应器工程国家重点实验室,上海200237;[2]中国科学院上海有机化学研究所,上海200032;[3]华东理工大学精细化工系,上海200237
年份:2001
卷号:27
期号:4
起止页码:353
中文期刊名:华东理工大学学报(自然科学版)
外文期刊名:Journal of East China University of Science and Technology
收录:CSTPCD;;Scopus;北大核心:【北大核心2000】;CSCD:【CSCD2011_2012】;
基金:上海市教育委员会基金资助项目
语种:中文
中文关键词:寡核苷酸探针;亚磷酰胺;巯基臂;荧光标记;固相合成法;三苯甲基保护;lmol基因
外文关键词:fluorescent; oligonucleotide probe; phosphoramidite; sulphydryl group; synthesis
摘要:用三苯甲基巯基乙醇与氯化甲氧基吗啡啉亚磷酰胺连接得到一种亚磷酰胺试剂 ( III)。用该试剂通过固相合成仪合成了 5′末端巯烷基修饰的寡核苷酸 ,用 Ag+ 脱去巯基的三苯甲基保护后与 ω-碘己基修饰的荧光基团反应 ,得到相应的荧光标记寡核苷酸探针。设计的 lmb H筛选探针与质粒 DNA进行杂交 ,杂交结果验证了这种标记的实用性 ,灵敏度可达 1
The sulphydryl group of mercaptoethanol is protected by a trityl group and then reacted with methoxymorpholinochlorophosphite to give the corresponding phosphoramidite (III). The phosphoramidite has been used to introduce tritylmercapto ethyl group to the 5′ terminus of an oligonucleotide by using a solid phase method in a DNA synthysizer. After deprotection by Ag +, the mercaptoethyl group carried by the oligonucleotide is tagged with a fluorescent dye. Designed probe for screening lmb H gene hybrids with plasmids, the useful method is proved by the results. It could be detected at a limit of lpmol.
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