SEQUENCE HOMOLOGY REQUIREMENTS FOR INTERMOLECULAR RECOMBINATION IN MAMMALIAN-CELLS
SEQUENCE HOMOLOGY REQUIREMENTS FOR INTERMOLECULAR RECOMBINATION IN MAMMALIAN-CELLS
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DOI：
10.1073/pnas.83.14.5199
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发表时间：
1986-07-01
影响因子：
11.1
通讯作者：
KUCHERLAPATI, R
中科院分区：
文献类型：
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作者：
AYARES, D;CHEKURI, L;KUCHERLAPATI, R
我们研究了导入人、猴和细菌细胞的质粒间分子间重组的同源性要求。构建了原核-真核穿梭载体pSV2neo的可变大小缺失衍生物。将这些质粒中的每一个与另一个含有不同的、非重叠缺失的pSV2neo质粒混合。哺乳动物细胞和细菌的重组是通过重建完整的neo基因的频率来衡量的。我们观察到25个碱基对的同源序列足以产生重组产物，这意味着在这种同源性水平下可以发生突触和同源配对。对产物的检测表明，非互惠重组在正常neo基因的产生中发挥了作用。此外，还观察到连锁标记的共转化。核外溶解作用似乎在基因转化中发挥了作用。
We have examined the homology requirements for intermolecular recombination between plasmids introduced into human, monkey, and bacterial cells. Variable-size-deletion derivatives of the prokaryotic-eukaryotic shuttle vector pSV2neo were constructed. Each of these plasmids was mixed with another pSV2neo plasmid containing a different, nonoverlapping deletion. Recombination was measured in mammalian cells and bacteria by the frequency of reconstruction of an intact neo gene. We observed that 25 base pairs of homologous sequence is sufficient to yield recombinant products, implying that synapsis and homologous pairing can occur with this level of homology. Examination of the products revealed that nonreciprocal recombination played a role in the generation of normal neo genes. In addition coconversion of linked markers was observed. Exonucleolytic action seems to play a role in gene conversion.