BIOMAT Database Logo BIOMAT Database Logo

Efficient secretion of biologically active recombinant OB protein (leptin) in Escherichia coli, purification from the periplasm and characterization. 1998

Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Roche Research Gent, Belgium.

The genes encoding the mature forms of mouse (mOB) and human OB (hOB) protein (also called leptin) were fused to the secretion signal coding sequence of the Escherichia coli outer membrane protein A (sOMP A). The hybrid genes were preceded by a ribosome binding site (RBS) and were expressed under transcriptional control of both the lipoprotein promoter (Plpp) and the lac promoter-operator (POlac). The recombinant fusion proteins were efficiently expressed and exported into the periplasmic compartment of E. coli cells from where they were recovered by osmotic shock as soluble mature polypeptides with the sOMP A precisely removed. Recombinant mOB and hOB proteins were also produced in Sf9 insect cells using the baculovirus expression system. Milligram quantities of both proteins were purified to homogeneity using ion-exchange, hydrophobic interaction chromatography and gel filtration and were found to be biologically active and to have antiobesity effects upon testing in genetically obese ob/ob mice.

UI MeSH Term Description Entries
D007274 Injections, Intraperitoneal Forceful administration into the peritoneal cavity of liquid medication, nutrient, or other fluid through a hollow needle piercing the abdominal wall. Intraperitoneal Injections,Injection, Intraperitoneal,Intraperitoneal Injection
D007276 Injections, Intraventricular Injections into the cerebral ventricles. Intraventricular Injections,Injection, Intraventricular,Intraventricular Injection
D008820 Mice, Obese Mutant mice exhibiting a marked obesity coupled with overeating, hyperglycemia, hyperinsulinemia, marked insulin resistance, and infertility when in a homozygous state. They may be inbred or hybrid. Hyperglycemic Mice,Obese Mice,Mouse, Hyperglycemic,Mouse, Obese,Hyperglycemic Mouse,Mice, Hyperglycemic,Obese Mouse
D009765 Obesity A status with BODY WEIGHT that is grossly above the recommended standards, usually due to accumulation of excess FATS in the body. The standards may vary with age, sex, genetic or cultural background. In the BODY MASS INDEX, a BMI greater than 30.0 kg/m2 is considered obese, and a BMI greater than 40.0 kg/m2 is considered morbidly obese (MORBID OBESITY).
D011506 Proteins Linear POLYPEPTIDES that are synthesized on RIBOSOMES and may be further modified, crosslinked, cleaved, or assembled into complex proteins with several subunits. The specific sequence of AMINO ACIDS determines the shape the polypeptide will take, during PROTEIN FOLDING, and the function of the protein. Gene Products, Protein,Gene Proteins,Protein,Protein Gene Products,Proteins, Gene
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D001835 Body Weight The mass or quantity of heaviness of an individual. It is expressed by units of pounds or kilograms. Body Weights,Weight, Body,Weights, Body
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D002850 Chromatography, Gel Chromatography on non-ionic gels without regard to the mechanism of solute discrimination. Chromatography, Exclusion,Chromatography, Gel Permeation,Chromatography, Molecular Sieve,Gel Filtration,Gel Filtration Chromatography,Chromatography, Size Exclusion,Exclusion Chromatography,Gel Chromatography,Gel Permeation Chromatography,Molecular Sieve Chromatography,Chromatography, Gel Filtration,Exclusion Chromatography, Size,Filtration Chromatography, Gel,Filtration, Gel,Sieve Chromatography, Molecular,Size Exclusion Chromatography
D002852 Chromatography, Ion Exchange Separation technique in which the stationary phase consists of ion exchange resins. The resins contain loosely held small ions that easily exchange places with other small ions of like charge present in solutions washed over the resins. Chromatography, Ion-Exchange,Ion-Exchange Chromatography,Chromatographies, Ion Exchange,Chromatographies, Ion-Exchange,Ion Exchange Chromatographies,Ion Exchange Chromatography,Ion-Exchange Chromatographies

Related Publications

Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Recombinant expression of biologically active rat leptin in Escherichia coli.
June 2001, Protein expression and purification,
Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Purification and characterization of biologically active recombinant human Eppin expressed in Escherichia coli.
April 2008, Chinese medical journal,
Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Posttranslational Targeting of a Recombinant Protein Promotes Its Efficient Secretion into the Escherichia coli Periplasm.
July 2019, Applied and environmental microbiology,
Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Efficient expression, purification and characterization of native human cystatin C in Escherichia coli periplasm.
July 2015, Protein expression and purification,
Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Secretion of active truncated CD4 into Escherichia coli periplasm.
April 1991, Applied microbiology and biotechnology,
Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Purification of biologically active human immunodeficiency virus rev protein from Escherichia coli.
November 1989, Virology,
Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Single-step purification of recombinant anthrax lethal factor from periplasm of Escherichia coli.
November 2006, Journal of biotechnology,
Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Highly efficient soluble expression, purification and characterization of recombinant Aβ42 from Escherichia coli.
May 2018, RSC advances,
Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Expression and purification of biologically active recombinant rabbit monocyte chemoattractant protein1 in Escherichia coli.
May 2018, FEMS microbiology letters,
Y Guisez, and I Faché, and L A Campfield, and F J Smith, and A Farid, and G Plaetinck, and J Van der Heyden, and J Tavernier, and W Fiers, and P Burn, and R Devos
Expression and purification of biologically active recombinant human paraoxonase 1 from inclusion bodies of Escherichia coli.
November 2015, Protein expression and purification,
Copied contents to your clipboard!