Abstract:Objective Construction of recombinant Bacillus subtilis for secreted expression of the porcine urate oxidase (pUOX). Methods pUOX-encoding gene pUOX was placed under the control of the P43 promoter and Bacillus subtilis signal peptide-encoding sequence.The pUOX was connected with a nprB signal peptide by SOE-PCR, then imported the shuttle vector pP43X. High-level expression and secretion of mature, authentic, and stable pUOX were achieved using the protease-deficient strain B. subtilis WB800 as the host. Results SDS-PAGE detected shows a clear protein band of about 34 kD, which is consistent with the expected molecular weight. Determination of enzyme activity of the recombinant protein is 2.322 U/ml. Conclusions Construction of recombinant Bacillus subtilis for secreted expression of the porcine urate oxidase is successful. Porcine urate oxisase can be expressed with a high activity.
李辉, 陈思, 马娇颖, 戴君, 谢光蓉, 陈建华. 重组枯草芽孢杆菌猪尿酸氧化酶基因工程菌的构建和分泌表达[J]. 《中国医药生物技术》杂志, 2012, 7(3): 187-190.
LI Hui, CHEN Si, MA Jiao-Ying, DAI Jun, XIE Guang-Rong, CHEN Jian-Hua. Construction of recombinant Bacillus subtilis for secreted expression of the porcine urate oxidase. , 2012, 7(3): 187-190.