Cloning and expression of the gene encoding the soluble cytochrome b562 of Escherichia coli.
Cloning and expression of the gene encoding the soluble cytochrome b562 of Escherichia coli.
复制标题
编码大肠杆菌可溶性细胞色素b562的基因的克隆和表达。
DOI：
10.1111/j.1432-1033.1991.tb16377.x
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Sligar,SG
中科院分区：
文献类型：
--
作者：
Nikkila,H;Gennis,RB;Sligar,SG
大肠杆菌可溶性细胞色素b562基因已克隆到aSalI片段上。对该基因的分析显示，在编码成熟蛋白的序列前面存在一个先导序列。使用ac启动子表达cytochromeb562产生的蛋白含量为总蛋白的3-5%。这种过度生产使得采用简单、高产的纯化方案获得均匀的细胞色素562成为可能。对纯化蛋白的光谱和N端序列分析表明，该蛋白与从me中纯化和鉴定的染色体表达的cytochromeb562相同。[J] Itagaki， E. & Hager， L. P.（1966）。医学杂志。化学通报，2004,22(3):387 - 395。研究表明，基因组序列编码了一个经典的N端信号序列，成熟的细胞色素b562被转移到质周空间。
The gene for the soluble cytochromeb562fromEscherichia coliB has been cloned on aSalI fragment. The analysis of the gene reveals the presence of a leader sequence in front of the sequence encoding the mature protein. Expression of cytochromeb562using thelac‐promoter produced the protein to a level of 3–5% of total protein. This over‐production enables employment of a simple, high‐yield purification protocol to obtain homogeneous cytochromeb562. Spectroscopic and N‐terminal sequence analyses of the purified protein demonstrate that it is identical to the chromosomally expressed cytochromeb562purified and characterized fromE. coliB [Itagaki, E. & Hager, L. P. (1966)J. Biol. Chem. 241, 3687–3695]. It is demonstrated that the genomic sequence codes for a classic N‐terminal signal sequence and that mature cytochromeb562is translocated to the periplasmic space.