A novel recombinant expression and purification approach for the full-length anti-apoptotic membrane protein Bcl-2.

abstract：:We examined the feasibility of high-level production of recombinant human prolactin, a multifunctional protein hormone, in insect cells using a baculovirus expression system. The human prolactin cDNA with and without the secretory signal sequence was cloned into pFastBac1 baculovirus vector under the control of polyhe...

journal_title：Protein expression and purification

authors： Das T,Johns PW,Goffin V,Kelly P,Kelder B,Kopchick J,Buxton K,Mukerji P

更新日期：2000-11-01 00:00:00

abstract：:Leptin is a 16-kDa nonglycosylated hormone that is produced in mature adipocytes and which acts primarily in the hypothalamus to reduce food intake and body weight. While the rat is a representative laboratory animal model in obesity research, so far recombinant rat leptin was not available. In the present study, rat ...

journal_title：Protein expression and purification

authors： Park JH,Lee HH,Na SY,Ju SK,Lee YJ,Lee MK,Kim KL

更新日期：2001-06-01 00:00:00

abstract：:The gene encoding a novel glucoamylase (GlucaM) from the Corallococcus sp. strain EGB was cloned and heterologous expressed in Escherichia coli BL21(DE3), and the enzymatic characterization of recombinant GlucaM (rGlucaM) was determined in the study. The glucaM had an open reading frame of 1938 bp encoding GlucaM of 6...

journal_title：Protein expression and purification

authors： Li Z,Ji K,Dong W,Ye X,Wu J,Zhou J,Wang F,Chen Q,Fu L,Li S,Huang Y,Cui Z

更新日期：2017-01-01 00:00:00

abstract：:This article reviews the development of immobilized-metal affinity chromatography (IMAC) and describes its most important applications. We provide an overview on the use of IMAC in protein fractionation and proteomics, in protein immobilization and detection, and on some special applications such as purification of im...

journal_title：Protein expression and purification

authors： Block H,Maertens B,Spriestersbach A,Brinker N,Kubicek J,Fabis R,Labahn J,Schäfer F

更新日期：2011-09-03 00:00:00

abstract：:3-Deoxy-d-manno-octulosonate 8-phosphate synthase (KDO8PS) [EC 4.1.2.16] is the first and rate-limiting enzyme in the 3-deoxy-d-manno-octulosonate (KDO) biosynthetic pathway. The enzyme is widely expressed in bacteria and plants. Their well conserved protein sequences imply a similar oligomeric arrangement. However, t...

journal_title：Protein expression and purification

authors： Zhang F,Xu Y,Zhang Z

更新日期：2014-09-01 00:00:00

abstract：:A hexahistidine tag was fused to the N-terminus of apoaequorin. A suitable vector encoding the fusion protein was constructed and used for transformation of Escherichia coli JM109 cells. Apoaequorin was overexpressed under the control of tac promoter. It was found, however, that most of the protein existed in the form...

journal_title：Protein expression and purification

authors： Glynou K,Ioannou PC,Christopoulos TK

更新日期：2003-02-01 00:00:00

abstract：:Interferons (IFNs) are a family of pleiotropic cytokines used for the treatment of various viral infections and cancers. The low-cost production of IFNs with high biological value and the discovery of IFNs with improved properties are important for the treatment of these diseases as well as for understanding the physi...

journal_title：Protein expression and purification

authors： Platis D,Foster GR

更新日期：2003-10-01 00:00:00

abstract：:Notch signaling plays a key role in cell differentiation and is very well conserved from Drosophila to humans. Ligands of Notch receptors are type I, membrane spanning proteins composed of a large extracellular region and a 100-150 residue cytoplasmic tail. We report here, for the first time, the expression, purificat...

journal_title：Protein expression and purification

authors： Popovic M,Coglievina M,Guarnaccia C,Verdone G,Esposito G,Pintar A,Pongor S

更新日期：2006-06-01 00:00:00

abstract：:A recombinant H(C) fragment of botulinum neurotoxin, serotype A (rBoNTA(H(C))), has been successfully expressed in a Mut(+) strain of the methylotrophic yeast Pichia pastoris for use as an antigen in a proposed human vaccine. Fermentation employed glycerol batch, glycerol-fed batch, and methanol-fed batch phases to ac...

journal_title：Protein expression and purification

authors： Potter KJ,Zhang W,Smith LA,Meagher MM

更新日期：2000-08-01 00:00:00

abstract：:The type I antifreeze proteins are simple amphipathic helical proteins found in abundance in polar fish species, where they act to prevent freezing of internal fluids by a mechanism of noncolligative freezing point depression. Large-scale production of these proteins for research and biotechnological purposes has been...

journal_title：Protein expression and purification

authors： Solomon RG,Appels R

更新日期：1999-06-01 00:00:00

abstract：:A mammalian expression vector with features optimized for simple expression and purification of secreted proteins has been developed. This vector was constructed to facilitate X-ray crystallographic studies of cysteine-rich glycoproteins that are difficult to express by other means. Proteins expressed with this vector...

journal_title：Protein expression and purification

authors： Leahy DJ,Dann CE 3rd,Longo P,Perman B,Ramyar KX

更新日期：2000-12-01 00:00:00

abstract：:The promyelocytic leukemia (PML) gene is involved in the 15/17 chromosomal translocation of acute promyelocytic leukemia (APL). It encodes a nuclear phosphoprotein containing an alpha-helical coiled-coil domain with four heptad repeats. The heptad repeats consist of four clusters of hydrophobic amino acids that mediat...

journal_title：Protein expression and purification

authors： Antolini F,Lo Bello M,Sette M

更新日期：2003-05-01 00:00:00

abstract：:Disulfide bonds are normally formed after a polypeptide has been exported from the reducing environment of the cytoplasm into a more oxidizing compartment, such as the bacterial periplasm. Recently, we showed that in Escherichia coli trxB gor mutants, in which the reduction of thioredoxin and glutathione is impaired, ...

journal_title：Protein expression and purification

authors： Levy R,Weiss R,Chen G,Iverson BL,Georgiou G

更新日期：2001-11-01 00:00:00

abstract：:Cell extracts of Clostridium kluyveri grown on ethanol plus succinate contained a NAD(H) dependent 4-hydroxybutanoate dehydrogenase (EC 1.1.1.61) at 66 U/mg. This enzyme was purified 42-fold under anaerobic conditions to homogeneity. Heat treatment, ion exchange chromatography on DEAE-cellulose, nondenaturing polyacry...

journal_title：Protein expression and purification

authors： Wolff RA,Kenealy WR

更新日期：1995-04-01 00:00:00

abstract：:Post-translational processing of host defense cathelicidin peptide LL-37 in human sweat and skin generates new antimicrobial peptides. To understand structure and mechanism of action of these LL-37 derivatives, this article presents the cloning and expression of SK-29, KR-20, LL-29, and LL-23. We also provide a two-st...

journal_title：Protein expression and purification

authors： Li Y,Li X,Wang G

更新日期：2007-10-01 00:00:00

abstract：:Influenza A virus displays one of the highest infection rates of all human viruses and therefore represents a severe human health threat associated with an important economical challenge. Influenza matrix protein 2 (M2) is a membrane protein of the viral envelope that forms a proton selective ion channel. Here we repo...

journal_title：Protein expression and purification

authors： Desuzinges Mandon E,Traversier A,Champagne A,Benier L,Audebert S,Balme S,Dejean E,Rosa Calatrava M,Jawhari A

更新日期：2017-03-01 00:00:00

abstract：:Strategies for the expression of precursors of eukaryotic secreted proteins as part of fused proteins in Escherichia coli have been explored. A fusion protein with beta-galactosidase at the N-terminal end and honeybee prepromelittin at the C-terminal end (beta-gal-pM) was expressed in low amounts as a cleaved polypept...

journal_title：Protein expression and purification

authors： He M,Adcock I,Chapman D,Lucy J,Austen B

更新日期：1991-10-01 00:00:00

abstract：:Pectate lyase (EC 4.2.2.2) gene from Bacillus subtilis RCK was cloned and expressed in Escherichia coli to maximize its production. In addition to soluble fraction, bioactive pectate lyase was also obtained from inclusion body aggregates by urea solubilization and refolding under in vitro conditions. Enzyme with speci...

journal_title：Protein expression and purification

authors： Kumar S,Jain KK,Singh A,Panda AK,Kuhad RC

更新日期：2015-06-01 00:00:00

abstract：:Streptavidin is a versatile molecule for many in vitro and in vivo applications. To optimize the production of the full-length streptavidin in a soluble and functional form via secretion using Bacillus subtilis as the expression host, three different strategies were used. These strategies include the construction of a...

journal_title：Protein expression and purification

authors： Wu SC,Hassan Qureshi M,Wong SL

更新日期：2002-04-01 00:00:00

abstract：:The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-t...

journal_title：Protein expression and purification

authors： Rosenlöcher J,Weilandt C,Sandig G,Reinke SO,Blanchard V,Hinderlich S

更新日期：2015-11-01 00:00:00

abstract：:The preparation of a pure and homogeneous protein sample at proper concentration is a prerequisite for success when attempting their crystallization for structural determination. The detergents suitable for solubilization particularly of membrane proteins are not always the best for crystallization. Myelin of the peri...

journal_title：Protein expression and purification

authors： Sedzik J,Uyemura K,Tsukihara T

更新日期：2002-12-01 00:00:00

abstract：:The methylotrophic yeasts Pichia pastoris and Pichia angusta (Hansenula polymorpha) were used for the comparative heterologous production of two model mammalian proteins of pharmaceutical interest, the NK1-fragment (22 kDa) of human hepatocyte growth factor and the extracellular domain (28 kDa) of mouse tissue factor ...

journal_title：Protein expression and purification

authors： Mack M,Wannemacher M,Hobl B,Pietschmann P,Hock B

更新日期：2009-08-01 00:00:00

abstract：:Pseudomonas fluorescens is a robust protein expression system that is very well suited for high throughput protein expression for structural genomics studies. Since NMR spectroscopy and X-ray crystallography are both used by various investigators in structure elucidation studies, the availability of target proteins la...

journal_title：Protein expression and purification

authors： Madduri K,Badger M,Li ZS,Xu X,Thornburgh S,Evans S,Dhadialla TS

更新日期：2009-05-01 00:00:00

abstract：:l-glutamate oxidase (GLOD), encoded by the gox gene, catalyses the transformation of l-glutamic acid into α-ketoglutaric acid (α-KG). In the present study, Pichia pastoris was used for heterologous production of GLOD following optimization of the gox coding sequence for expression in the yeast host. A series of constr...

journal_title：Protein expression and purification

authors： YaPing W,Ben R,Hong Y,Rui H,Li L,Ping'an L,Lixin M

更新日期：2017-01-01 00:00:00

abstract：:A gene encoding a glutamate-specific endopeptidase (GSE) from Bacillus licheniformis (BL) has been cloned in Escherichia coli cells. The recombinant protein was expressed as cytoplasmic insoluble inclusion bodies. Immobilized metal affinity chromatography was employed to purify the protein, and then a 27-kDa GSE inter...

journal_title：Protein expression and purification

authors： Ye W,Liu J,Wang H,Wang J,Wang X

更新日期：2012-03-01 00:00:00

abstract：:Structural studies of human peptide hormone somatostatin 14 (SS14) require high amounts of isotopically labelled SS14 to be produced. Here we report a method for effective production of isotopically labelled SS14. SS14 was expressed as a fusion protein with thioredoxin in Escherichia coli. Co-expression of a longer po...

journal_title：Protein expression and purification

authors： Nespovitaya N,Barylyuk K,Eichmann C,Zenobi R,Riek R

更新日期：2014-07-01 00:00:00

abstract：:Triacylglycerol hydrolase mobilizes stored triacylglycerol some of which is used for very-low-density lipoprotein assembly in the liver. A full-length cDNA coding for a human triacylglycerol hydrolase (hTGH) was isolated from a human liver cDNA library. The cDNA has an open reading frame of 576 amino acids with a clea...

journal_title：Protein expression and purification

authors： Alam M,Ho S,Vance DE,Lehner R

更新日期：2002-02-01 00:00:00

abstract：:An active form of a single-chain antibody fragment (scFv) from the murine monoclonal antibody ABL-1, which is specific for B-cell-activating factor of the TNF family, was produced in Escherichia coli. The complementary DNAs encoding the variable regions of the heavy chain (VH) and light chain (VL) were connected by a ...

journal_title：Protein expression and purification

authors： Cao P,Tang XM,Guan ZB,Diao ZY,Zhang SQ

更新日期：2005-10-01 00:00:00

abstract：:The erm proteins confer resistance to the MLS (macrolide-lincosamide-streptogramin B) antibiotics in various microorganisms, including pathogens, through dimethylation of a single adenine residue (A2085: Bacillus subtilis coordinate) of the 23S rRNA to reduce the affinity of antibiotics, thereby enabling the cells to ...

journal_title：Protein expression and purification

authors： Jin HJ,Yang YD

更新日期：2002-06-01 00:00:00

abstract：:The HIV-1 Gag precursor protein, Pr55(Gag), is a multi-domain polyprotein that drives HIV-1 assembly. The morphological features of HIV-1 suggested Pr55(Gag) assumes a variety of different conformations during virion assembly and maturation, yet structural determination of HIV-1 Pr55(Gag) has not been possible due to ...

journal_title：Protein expression and purification

authors： McKinstry WJ,Hijnen M,Tanwar HS,Sparrow LG,Nagarajan S,Pham ST,Mak J

更新日期：2014-08-01 00:00:00