Group II introns as controllable gene targeting vectors for genetic manipulation of bacteria
Group II introns as controllable gene targeting vectors for genetic manipulation of bacteria
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DOI：
10.1038/nbt1201-1162
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发表时间：
2001-12-01
影响因子：
46.9
通讯作者：
Lambowitz, AM
中科院分区：
文献类型：
--
作者：
Karberg, M;Guo, HT;Lambowitz, AM
可移动的第二组内含子可以被重定向以插入到几乎任何所需的DNA靶点。在这里，我们证明了重定向第二组内含子可以用于在大肠杆菌和其他细菌中以0.1%-22%的频率进行高度特异性的染色体基因破坏。此外，内含子可以用来引入有针对性的染色体断裂，这些断裂可以通过与同源DNA片段转化来修复，从而能够引入点突变。由于其广泛的宿主范围，可移动的第二组内含子应该在广泛的细菌中用于基因工程和功能基因组学。
Mobile group II introns can be retargeted to insert into virtually any desired DNA target. Here we show that retargeted group II introns can be used for highly specific chromosomal gene disruption in Escherichia coli and other bacteria at frequencies of 0.1-22%. Furthermore, the introns can be used to introduce targeted chromosomal breaks, which can be repaired by transformation with a homologous DNA fragment, enabling the introduction of point mutations. Because of their wide host range, mobile group II introns should be useful for genetic engineering and functional genomics in a wide variety of bacteria.