Center for Environmental Technology Research, Korea Institute of Science and Technology, Seoul, Republic of Korea.
Appl Biochem Biotechnol. 2010 May;161(1-8):75-83. doi: 10.1007/s12010-009-8788-4. Epub 2009 Oct 2.
Degeneration is one of the limiting factors in butanol fermentation, and it must be monitored and prevented for stable butanol production. In Clostridium acetobutylicum ATCC 824, the most well-known butanol-producing microorganism, degeneration is caused by the loss of the pSOL1 plasmid that carries essential genes involved in solvent production. In this study, we designed two specific primer and probe sets for real-time qPCR (RT-qPCR) detection of C. acetobutylicum ATCC 824 (the C. aceto set) and pSOL1-possessing C. acetobutylicum ATCC 824 (the DGS set). Specific primer and probe sets were designed on the basis of the 16S rDNA sequence and pSOL1 sequence. The number of degenerated C. acetobutylicum could be quantified by subtracting the number of C. acetobutylicum ATCC 824 containing pSOL1 from the total number of C. acetobutylicum ATCC 824. The primer and probe sets permitted the specific detection and quantification of degenerated C. acetobutylicum and total butanol-producing C. acetobutylicum by RT-qPCR.
退化是丁醇发酵的限制因素之一，必须对其进行监测和预防，以实现稳定的丁醇生产。在丙酮丁醇梭菌 ATCC 824 中，这种最知名的产丁醇微生物，退化是由于携带溶剂生产必需基因的 pSOL1 质粒的丢失引起的。在本研究中，我们设计了两个用于实时 qPCR (RT-qPCR) 检测丙酮丁醇梭菌 ATCC 824 的特异性引物和探针集（C. aceto 集）和携带 pSOL1 的丙酮丁醇梭菌 ATCC 824（DGS 集）。特异性引物和探针集是基于 16S rDNA 序列和 pSOL1 序列设计的。通过从丙酮丁醇梭菌 ATCC 824 的总数中减去含有 pSOL1 的丙酮丁醇梭菌 ATCC 824 的数量，可以定量计算退化的丙酮丁醇梭菌的数量。通过 RT-qPCR，该引物和探针集允许对退化的丙酮丁醇梭菌和总产丁醇的丙酮丁醇梭菌进行特异性检测和定量。
