CLONING AND EXPRESSION OF THE GENE ENCODING THE SOLUBLE CYTOCHROME-B562 OF ESCHERICHIA-COLI

被引：49

作者：

NIKKILA, H ^{[1
]}

GENNIS, RB ^{[1
]}

SLIGAR, SG ^{[1
]}

机构：

[1] UNIV ILLINOIS,DEPT BIOCHEM,1209 W CALIF ST,URBANA,IL 61801

来源：

EUROPEAN JOURNAL OF BIOCHEMISTRY | 1991年 / 202卷 / 02期

关键词：

D O I：

10.1111/j.1432-1033.1991.tb16377.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The gene for the soluble cytochrome b562 from Escherichia coli B has been cloned on a SalI fragment. The analysis of the gene reveals the presence of a leader sequence in front of the sequence encoding the mature protein. Expression of cytochrome b562 using the lac-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 cytochrome b562. Spectroscopic and N-terminal sequence analyses of the purified protein demonstrate that it is identical to the chromosomally expressed cytochrome b562 purified and characterized from E. coli B [Itagaki, E. & Hager, L. P. (1966) J. Biol. Chem. 241, 3687-36951. It is demonstrated that the genomic sequence codes for a classic N-terminal signal sequence and that mature cytochrome b562 is translocated to the periplasmic space.

引用

页码：309 / 313

页数：5

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