Green E M, Bennett G N
Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77251, USA.
Appl Biochem Biotechnol. 1996 Spring;57-58:213-21. doi: 10.1007/BF02941702.
A nonreplicative plasmid containing an internal aad gene fragment has been integrated into the chromosome of Clostridium acetobutylicum ATCC 824. Transformation was accomplished by electroporation with relatively high concentrations of methylated plasmid DNA. Southern hybridization experiments revealed that integration occurred by single crossover homologous recombination inactivating the aad gene. Integrants were relatively stable after 25 generations. Inactivation of the aad gene drastically reduced solvent production. This result suggests that aldehyde/alcohol dehydrogenase(AAD) plays a important role in butanol production.
一个含有内部aad基因片段的非复制性质粒已整合到丙酮丁醇梭菌ATCC 824的染色体中。通过用相对高浓度的甲基化质粒DNA进行电穿孔来完成转化。Southern杂交实验表明，整合是通过单交换同源重组使aad基因失活而发生的。整合体在25代后相对稳定。aad基因的失活极大地降低了溶剂产量。这一结果表明醛/醇脱氢酶（AAD）在丁醇生产中起重要作用。
