The characteristics of Grx4 and its interaction with ClpYQ protease in Escherichia coli

• Main Authors: Chun-Yang Chang, 張鈞暘
• Other Authors: 吳蕙芬
• Format: Others
• Language: zh-TW
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/yex447

博士 === 國立臺灣大學 === 農業化學研究所 === 107 === The glutaredoxin-4 (Grx4)-encoding in grxD gene in Escherichia coli, which is the only monothiol glutaredoxin (only active region with CGFS) found in it. In this study, grxD-lacZ fusion gene and Grx4-3xFLAG in chromosome were used to observe the regulation of grxD. When small RNA was expressed in a high level, FnrS negatively regulated grxD-lacZ to about 1/2 of the original expression, at the level of Grx4-3xFLAG protein the more obvious repression happend. RyhB also negatively regulates the performance of grxD-lacZ by ~20%. In addition, in the absence of transcription factors, NsrR was found to be involved in the positive regulation of grxD, and nsrR deletion caused a decrease in performance by nearly 1/2, while the Grx4-3xFLAG protein level was more influential. Therefore, the performance of grxD-lacZ is regulated by factors such as oxygen, iron, and nitric oxide related factors, and the influence of Grx4-3xFLAG on growth indicates the importance of fine regulation of this part. In addition to gene regulation studies, P1 phage plays an important role in E. coli genetic engineering and be used to construct these various mutants. Therefore, this paper also overcomes the problem of easy inactivation of P1 before, and utilizes new proliferation, the way to make P1 work well in genetic engineering operations. Finally, with the previous in-depth study of SulA and ClpYQ, we have developed a set of "new enhance cloning and transformation efficiency method and cell division inhibition technology" and expected to be applied in genetic engineering, microbial population analysis and other research.