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Concurrent targeted exchange of three bases in mammalian hprt by oligonucleotides.

Abstract：

:The repair of point mutations in hprt gene by single-stranded oligonucleotides represents a model to test targeted nucleotide exchange. We studied the concurrent nucleotide exchange of two or three nucleotides in the hprt deficient hamster cell line V79-151. The used oligonucleotides resulted in mismatches at two (151, 159) or three (151, 144, and 159) hprt positions. The hprt point mutation at position 151 was repaired in about 2/10(6) cells as shown by hprt sequencing in clones surviving HAT selection. The second nucleotide exchange at hprt position 159 was found in 7% of these HAT selected clones. Using oligonucleotides resulting in three mismatches, 29% of the clones showed nucleotide exchanges at the two hprt positions (151, 144) and about 4% at three positions (151, 144, and 159). These results indicate that single-stranded oligonucleotides can generate two or three nucleotide exchanges in a mammalian chromosomal gene.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Kenner O,Lutomska A,Speit G,Vogel W,Kaufmann D

doi

10.1016/j.bbrc.2004.07.062

subject

Has Abstract

pub_date

2004-09-03 00:00:00

pages

1017-23

issue

4

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(04)01560-8

journal_volume

321

pub_type

杂志文章

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