Kaneko Shinya, Tsuge Kenji, Takeuchi Takashi, Itaya Mitsuhiro
Mitsubishi Kagaku Institute of Life Science, 11 Minamiooya, Machida, Tokyo 194-8511, Japan.
Nucleic Acids Res. 2003 Sep 15;31(18):e112. doi: 10.1093/nar/gng114.
A novel genome vector using the 4215 kb Bacillus subtilis genome provides for precise target cloning and processing of the cloned DNA to the desired structure. Each process highly dependent on homologous recombination in the host B.subtilis is distinguished from the other cloning systems. A 120 kb mouse jumonji (jmj) genomic gene was processed in the genome vector to give a series of truncated sub-megasized DNA. One of these truncated segments containing the first intron was copied in a plasmid by a recombinational transfer method developed for B.subtilis. DNA manipulation previously considered difficult is argued with respect to DNA size and accuracy.
一种使用4215 kb枯草芽孢杆菌基因组的新型基因组载体可实现精确的靶标克隆,并将克隆的DNA加工成所需结构。每个过程高度依赖宿主枯草芽孢杆菌中的同源重组,这与其他克隆系统不同。一个120 kb的小鼠jumonji(jmj)基因组基因在基因组载体中进行加工,得到一系列截短的亚兆碱基大小的DNA。其中一个包含第一个内含子的截短片段通过为枯草芽孢杆菌开发的重组转移方法在质粒中进行复制。针对DNA大小和准确性,讨论了以前认为困难的DNA操作。