Yeast artificial chromosome segregation from host chromosomes with similar lengths.
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.
Izvolsky
K I
KI
Center for Advanced Biotechnology, Boston University, 36 Cummington Street, Boston, MA 02215, USA.
Demidov
V V
VV
Bukanov
N O
NO
Frank-Kamenetskii
M D
MD
D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D015825:Chromosomes, Fungal / Q000235:genetics*; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D016521:Electrophoresis, Gel, Pulsed-Field; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D020135:Peptide Nucleic Acids; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D004271:DNA, Fungal; D020135:Peptide Nucleic Acids