Olson Daniel G, Lynd Lee R
Thayer School of Engineering at Dartmouth College, Hanover, New Hampshire, USA.
Methods Enzymol. 2012;510:317-30. doi: 10.1016/B978-0-12-415931-0.00017-3.
In this work, we provide detailed instructions for transformation of Clostridium thermocellum by electroporation. In addition, we describe two schemes for genetic modification: allelic replacement-where the gene of interest is replaced by an antibiotic marker and markerless gene deletion-where the gene of interest is removed and the selective markers are recycled. The markerless gene deletion technique can also be used for insertion of genes onto the C. thermocellum chromosome.
在本研究中，我们提供了通过电穿孔法转化嗜热栖热菌的详细操作说明。此外，我们描述了两种基因改造方案：等位基因替换——将目标基因替换为抗生素标记；无标记基因缺失——去除目标基因并回收选择标记。无标记基因缺失技术也可用于将基因插入嗜热栖热菌染色体。
