Smolina Irina V, Kuhn Heiko, Lee Charles, Frank-Kamenetskii Maxim D
Center for Advanced Biotechnology and Department of Biomedical Engineering, Boston University, 36 Cummington Street, MA 02215, USA.
Bioorg Med Chem. 2008 Jan 1;16(1):84-93. doi: 10.1016/j.bmc.2007.04.063. Epub 2007 May 5.
The ability of peptide nucleic acid (PNA) to open up duplex DNA in a highly sequence-specific manner makes it possible to detect short DNA sequences on the background of or within genomic DNA under non-denaturing conditions. To do so, chosen marker sites in double-stranded DNA are locally opened by a pair of PNA openers, thus transforming one strand within the target region (20-30 bp) into the single-stranded form. Onto this accessible DNA sequence a circular oligonucleotide probe is assembled, which serves as a template for rolling circle amplification (RCA). Both homogeneous and heterogeneous assay formats are investigated, as are different formats for fluorescence-based amplicon detection. Our recent data with immobilized analytes suggest that marker sequences in plasmid and bacterial chromosomal DNA can be successfully detected.
肽核酸（PNA）以高度序列特异性方式打开双链DNA的能力，使得在非变性条件下，能够在基因组DNA背景中或内部检测短DNA序列。为此，双链DNA中选定的标记位点由一对PNA开启剂局部打开，从而将目标区域（20 - 30个碱基对）内的一条链转化为单链形式。在这个可及的DNA序列上组装一个环状寡核苷酸探针，它作为滚环扩增（RCA）的模板。研究了均相和非均相检测形式，以及基于荧光的扩增子检测的不同形式。我们最近关于固定化分析物的数据表明，质粒和细菌染色体DNA中的标记序列能够被成功检测。
