英语翻译摘 要在分子生物学研究中,DNA的序列分析是进一步研究和改造目的基因的基础,测序更是序列分析的必不可少的环节.当前应用最广泛的测序技术双脱氧链末端终止法.本研究涉及一种
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英语翻译摘 要在分子生物学研究中,DNA的序列分析是进一步研究和改造目的基因的基础,测序更是序列分析的必不可少的环节.当前应用最广泛的测序技术双脱氧链末端终止法.本研究涉及一种
英语翻译
摘 要
在分子生物学研究中,DNA的序列分析是进一步研究和改造目的基因的基础,测序更是序列分析的必不可少的环节.当前应用最广泛的测序技术双脱氧链末端终止法.本研究涉及一种称之为“alpha-thiosequencing”的新型DNA测序技术,该技术主要是利用了外切酶去除延伸上的荧光信号以及alpha硫代核苷酸保证延伸的进行.将扩增产物固定在固相基底上,变性去除互补链之后,杂交上硫代修饰的测序引物,用含有单一已知核苷酸单体混合物(荧光标记的与正常的核苷酸单体)的延伸反应溶液进行延伸反应形成核苷键,通过外切酶Ⅲ的3'→5'外切活性去除延伸上的荧光碱基;延伸具有相同类型的硫代核苷酸单体,使之形成硫代核苷键,拮抗下一次的酶切反应,确保引物进一步向前延伸,用含第二种已知核苷酸单体的延伸反应溶液进行延伸反应.根据碱基互补配对原则,只有加入的核苷酸单体与紧接着延伸的模板上的碱基相互配对,延伸反应才能进行,因此,四种碱基(A、T、C、G)依次轮流加入,最终就能模板序列的碱基信息.该方法将为人们生命过程的分子机制研究、基因组DNA序列分析、个体化医学、疾病的预测和预防、药物的开发和个体化研究等具有较大的应用价值.
关键词:核苷酸单体、荧光信号、碱基、测序
英语翻译摘 要在分子生物学研究中,DNA的序列分析是进一步研究和改造目的基因的基础,测序更是序列分析的必不可少的环节.当前应用最广泛的测序技术双脱氧链末端终止法.本研究涉及一种
In the research of molecular biology, The sequence analysis of DNA is On the basis of
further researching and reformming of destination gene sequencing , even Test sequence is the essential link of equence analysis . The most widely used sequencing is dideoxy chain termination method. This study relates to a new DNA sequencing technology called" alpha-thiosequencing" , the technology is mainly used to remove the exonuclease extending fluorescence signal and the fact that alpha nucleotides extends the thiosulfate. The amplification product is fixed on the solid substrate, removing the complementary strand degeneration after hybridization, the phosphorothioate modified sequencing primers, containing single known nucleotide monomer mixture ( fluorescently labeled with normal nucleotide monomers ) extension reaction solution is formed extending reaction nucleosides keys via exonuclease III, the3 ' - > 5' exonuclease removal of the fluorescent activity extends base; extension has the same type of thiosulfate nucleotide monomers, so that the formation of phosphorothioate nucleosides keys, antagonizes the next enzyme reaction, to ensure that the primer extends forward, with second known nucleotide monomer extension reaction solution extension reaction. According to the principle of complementary base pairing, only joining the nucleotide monomers and then extending the base template matching, extending the reaction can be carried out, therefore, the four base pairs ( A, T, C, G ) in turn, can eventually template sequence base information. The method will provide people the life course research, molecular mechanisms of genomic DNA sequence analysis, individualized medicine, disease prediction and prevention, drug development and individualized study has greater application value.