Ozawa Kazumichi, Harashina Takeyori, Yatsunami Rie, Nakamura Satoshi
Department of Bioengineering, Tokyo Institute of Technology, Yokohama, Japan.
Extremophiles. 2005 Aug;9(4):281-8. doi: 10.1007/s00792-005-0443-6. Epub 2005 Apr 21.
The gene encoding a cell division protein FtsZ1 was cloned from an extremely halophilic archaeon, Haloarcula japonica strain TR-1. Nucleotide sequencing analysis of the ftsZ1 gene revealed that the structural gene consisted of an open reading frame of 1,158 nucleotides encoding 386 amino acids. Transcription of the ftsZ1 gene in Ha. japonica was confirmed by RT-PCR. A modified ftsZ1 gene was inserted into the shuttle vector pWL102 and used to transform Ha. japonica. The recombinant FtsZ1 was produced as a fusion with hexahistidine-tag in Ha. japonica host cells and purified. Purified recombinant FtsZ1 exhibited GTP-dependent polymerization activity and GTP-hydrolyzing activity in the presence of high concentrations of KCl.
编码细胞分裂蛋白FtsZ1的基因是从极端嗜盐古菌日本嗜盐碱杆菌TR-1菌株中克隆得到的。ftsZ1基因的核苷酸测序分析表明，该结构基因由一个1158个核苷酸的开放阅读框组成，编码386个氨基酸。通过逆转录聚合酶链反应（RT-PCR）证实了ftsZ1基因在日本嗜盐碱杆菌中的转录。将一个修饰后的ftsZ1基因插入穿梭载体pWL102中，并用于转化日本嗜盐碱杆菌。重组FtsZ1在日本嗜盐碱杆菌宿主细胞中作为与六组氨酸标签融合的蛋白产生并进行了纯化。纯化后的重组FtsZ1在高浓度氯化钾存在的情况下表现出GTP依赖性聚合活性和GTP水解活性。
