Transformation with oligonucleotides creating clustered changes in the yeast genome.

Transformation with oligonucleotides creating clustered changes in the yeast genome.
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DOI:
10.1371/journal.pone.0042905
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发表时间:
2012
期刊:
影响因子:
3.7
通讯作者:
Crouse GF
Crouse GF
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Rodriguez GP;Song JB;Crouse GF

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We have studied single-strand oligonucleotide (oligo) transformation of yeast by using 40-nt long oligos that create multiple base changes to the yeast genome spread throughout the length of the oligos, making it possible to measure the portions of an oligo that are incorporated during transformation. Although the transformation process is greatly inhibited by DNA mismatch repair (MMR), the pattern of incorporation is essentially the same in the presence or absence of MMR, whether the oligo anneals to the leading or lagging strand of DNA replication, or whether phosphorothioate linkages are used at either end. A central core of approximately 15 nt is incorporated with a frequency of >90%; the ends are incorporated with a lower frequency, and loss of the two ends appears to be by different mechanisms. Bases that are 5–10 nt from the 5′ end are generally lost with a frequency of >95%, likely through a process involving flap excision. On the 3′ end, bases 5–10 nt from the 3′ end are lost about 1/3 of the time. These results indicate that oligos can be used to create multiple simultaneous changes to the yeast genome, even in the presence of MMR.
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