Cloning and nucleotide sequence of the gcv operon encoding the Escherichia coli glycine-cleavage system.
Cloning and nucleotide sequence of the gcv operon encoding the Escherichia coli glycine-cleavage system.
复制标题
编码大肠杆菌甘氨酸切割系统的 gcv 操纵子的克隆和核苷酸序列。
DOI：
10.1111/j.1432-1033.1993.tb18172.x
复制
发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Y. Motokawa
中科院分区：
文献类型：
--
作者：
K. Okamura;Yosuke Ohmura;K. Fujiwara;Y. Motokawa
从大肠杆菌中纯化了甘氨酸裂解体系的p蛋白、h蛋白和t蛋白。测定了它们的n端氨基酸序列，设计了一套用于基因克隆的寡核苷酸探针。大肠杆菌染色体62分钟区域附近的DNA片段的核苷酸序列，包含甘氨酸-切割系统成分的基因已经确定。该序列包含3个结构基因，分别编码t蛋白（363个氨基酸，40013 Da）、h蛋白（128个氨基酸，13679 Da）和p蛋白（956个氨基酸，104240 Da）。这些基因分别被命名为gcvT、gcvH和gcvP。它们以上述顺序排列在同一条DNA链上，具有短的顺反子间序列。gcvT之前存在一个潜在的启动子，gcvP之后存在一个典型的rho独立终止子序列，表明这三个基因构成了一个单一的操纵子。大肠杆菌甘氨酸-裂解系统的每个组成部分与动物和植物的氨基酸序列相当相似。将含有gcv操纵子的质粒转染到大肠杆菌细胞中，gcvT、gcvH和gcvP的基因产物在gcv操纵子启动子的引导下过表达。然而，携带含有gcv操纵子而不含启动子区域和gcvT的5′端部分的质粒的细菌未能过表达这三种成分中的任何一种。
P-protein, H-protein and T-protein of the glycine cleavage system have been purified from Escherichia coli. Their N-terminal amino acid sequences were determined, and a set of oligonucleotide probes was designed for gene cloning. The nucleotide sequence of a fragment of DNA around the 62-min region of the E. coli chromosome, containing genes for the components of the glycine-cleavage system has been determined. The sequence includes three structural genes encoding T-protein (363 amino acids, 40013 Da), H-protein (128 amino acids, 13679 Da) and P-protein (956 amino acids, 104240 Da). These genes are named gcvT, gcvH and gcvP, respectively. They are organized in the above-mentioned order on the same strand of DNA with short intercistronic sequences. The presence of a potential promoter preceding gcvT and a typical rho-independent terminator sequence following gcvP indicated that the three genes constitute a single operon. Each component of the E. coli glycine-cleavage system exhibits considerable amino acid sequence similarity with the animal and plant counterparts. When the plasmid containing the gcv operon was transfected in E. coli cells, the gene products of gcvT, gcvH and gcvP were overexpressed under the direction of the promoter of the gcv operon. However, bacteria harboring the plasmid that contained the gcv operon without the promoter region and the 5' terminal portion of gcvT failed to overexpress any of the three components.
一种大肠杆菌蛋白，与豌豆和鸡肝中的甘氨酸裂解酶复合物的 H 蛋白具有同源性。
DOI：
10.3109/10425179109008434
发表时间：
1991
期刊：
DNA sequence : the journal of DNA sequencing and mapping
影响因子：
--
作者：
Stauffer,LT;Steiert,PS;Steiert,JG;Stauffer,GV
通讯作者：
Stauffer,GV