Sinumvayo Jean Paul, Yang Sen, Chen Jian, Du Guocheng, Kang Zhen
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi 214122, Jiangsu, China.
Sheng Wu Gong Cheng Xue Bao. 2017 Jul 25;33(7):1091-1100. doi: 10.13345/j.cjb.160484.
Terminators as regulatory signals are typically placed behind the last coding sequence to block the transcription of DNA to RNA and release the transcript. In the present study, the hairpin and the U-rich sequence of the bacteriophage λto terminator were first modified to investigate their effects on termination efficiency and mRNA stability in Bacillus subtilis 168. Compared with the native λto terminator, the terminator variants M3, M11 and M12 showed higher termination efficiency values. Moreover, the variantsM3, M4 and M11 showed significant positive effects on the mRNA stability of the upstream gfp gene. Additionally, insertion of RNase site also increased the mRNA stability. The results of this study suggested that the composition of the hairpin loop is not required for effective intrinsic termination in B. subtilis. Our results also showed that the terminator could also be used as a potential tool for increasing mRNA stability and the corresponding enzyme production in B. subtilis.
作为调控信号的终止子通常置于最后一个编码序列之后,以阻断DNA转录为RNA并释放转录本。在本研究中,首先对噬菌体λto终止子的发夹结构和富含U的序列进行修饰,以研究它们对枯草芽孢杆菌168终止效率和mRNA稳定性的影响。与天然λto终止子相比,终止子变体M3、M11和M12表现出更高的终止效率值。此外,变体M3、M4和M11对上游gfp基因的mRNA稳定性具有显著的正向影响。另外,插入核糖核酸酶位点也提高了mRNA稳定性。本研究结果表明,发夹环的组成并非枯草芽孢杆菌有效固有终止所必需。我们的结果还表明,该终止子还可作为提高枯草芽孢杆菌mRNA稳定性及相应酶产量的潜在工具。