CODING PROPERTIES OF A UNIQUE APURINIC APYRIMIDINIC SITE REPLICATED IN MAMMALIAN-CELLS
CODING PROPERTIES OF A UNIQUE APURINIC APYRIMIDINIC SITE REPLICATED IN MAMMALIAN-CELLS
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DOI：
10.1006/jmbi.1994.1457
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发表时间：
1994-07-29
影响因子：
5.6
通讯作者：
GENTIL, A
中科院分区：
文献类型：
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作者：
NETO, JBC;CABRAL, REC;GENTIL, A
利用携带独特基本位点的单链穿梭载体，在哺乳动物细胞中研究了基本位点的编码特性。该探针是由插入到单链载体中的含有尿嘧啶的寡核苷酸用尿嘧啶DNA糖基酶消化而成的。在能够支持DNA复制的猴COS7细胞中复制该载体后，在细菌中分离出该质粒后代。对回收的质粒进行DNA测序表明，基本位点的复制不会导致与非编码位点相反的给定碱基的优先插入。这四个碱基的插入频率与随机分布没有统计学上的不同。因此，对于在哺乳动物细胞中复制的外源单链DNA，与唯一的基本位点相反的“A规则插入”似乎不适用，至少对我们使用的序列不适用。为了复制基本位点，不需要通过宿主细胞的预处理来诱导类似SOS的条件。
The coding properties of abasic sites have been studied in mammalian cells using a single-stranded shuttle vector carrying a unique abasic site. The probe was produced by digestion with the uracil DNA glycosylase of a uracil-containing oligonucleotide which was inserted in the single-stranded vector. After replication in monkey COS7 cells able to support DNA replication of the vector, the plasmid progeny were isolated in bacteria. DNA sequencing of rescued plasmids showed that replication of abasic sites does not lead to preferential insertion of a given base opposite the non-coding site. The four bases were inserted with a frequency which was not statistically different from a random distribution. It appears therefore that the "A rule insertion" opposite a unique abasic site does not apply, at least with the sequence we used, for an exogeneous single-stranded DNA replicated in mammalian cells. It was not necessary to induce SOS-like conditions by pretreatment of host cells, in order to replicate abasic sites.