Izvolsky K I, Demidov V V, Bukanov N O, Frank-Kamenetskii M D
Center for Advanced Biotechnology, Boston University, 36 Cummington Street, Boston, MA 02215, USA.
Nucleic Acids Res. 1998 Nov 1;26(21):5011-2. doi: 10.1093/nar/26.21.5011.
We propose a new method for segregation of yeast artificial chromosomes (YACs) from endogenous yeast chromosomes with similar lengths. The method is based on recently developed PNA-assisted rare cleavage (PARC) of genomic DNA. We apply the PARC procedure to YAC-containing samples of yeast DNA in such a way that host chromosomes, which electrophoretically comigrate with the chosen YACs, are selectively digested while YACs remain intact. These data demonstrate that a pool of appropriate PNAs can be used as an efficient tool for the PARC-based isolation of intact purified YACs directly from the host cells.
我们提出了一种从长度相似的内源性酵母染色体中分离酵母人工染色体(YAC)的新方法。该方法基于最近开发的基因组DNA的肽核酸(PNA)辅助稀有切割(PARC)。我们将PARC程序应用于含YAC的酵母DNA样本,使得与所选YAC在电泳中迁移率相同的宿主染色体被选择性消化,而YAC保持完整。这些数据表明,一组合适的PNA可作为一种有效的工具,用于直接从宿主细胞中基于PARC分离完整纯化的YAC。