Cloning, purification, and crystallization of a bacterial gene expression regulator--Hfq protein from Escherichia coli.
Thermostable RNA-binding protein Hfq (also denoted HF1) is a multifunctional expression regulator of many bacterial genes. The regulation takes place both at a translation level (directly) and transcription level (indirectly through the stimulation of bacterial RNA polymerase sigmaS-subunit translation). We have cloned and overexpressed the hfq gene from E. coli and developed a purification procedure for the protein. Using gel filtration and ultracentrifugation techniques it was shown that the obtained Hfq protein is highly homogeneous and well dissolved. It has been crystallized and can be used for structural investigations.
Vassilieva
I M
IM
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, 142290, Russia. yuvas@vega.protres.ru
Rouzanov
M V
MV
Zelinskaya
N V
NV
Moll
I
I
Bläsi
U
U
Garber
M B
MB
D000595:Amino Acid Sequence; D002850:Chromatography, Gel / Q000379:methods; D003001:Cloning, Molecular; D003460:Crystallization; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics; D035001:Host Factor 1 Protein / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016601:RNA-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D014461:Ultracentrifugation / Q000379:methods
D017931:DNA Primers; D029968:Escherichia coli Proteins; C087373:Hfq protein, E coli; D035001:Host Factor 1 Protein; D016601:RNA-Binding Proteins