A screening strategy for heterologous protein expression in Escherichia coli with the highest return of investment.

abstract：:Glycolate oxidase is a flavin-dependent enzyme in the photorespiratory pathway in plants. Here we report the heterologous expression of glycolate oxidase in Escherichia coli and an isolation procedure which results in 4 mg pure protein per gram cell paste in only 1.5 days. This corresponds to a more than 50-fold impro...

journal_title：Protein expression and purification

authors： Stenberg K,Lindqvist Y

更新日期：1996-11-01 00:00:00

abstract：:Production of VCSM13 phage displaying a high density of CD147 ectodomain (CD147Ex) was achieved when culturing conditions were modulated. A phagemid expressing CD147Ex was constructed and used to produce phage display CD147Ex gpVIII fusion protein in TG1 Escherichia coli. Displaying of CD147Ex via gpVIII was successfu...

journal_title：Protein expression and purification

authors： Intasai N,Arooncharus P,Kasinrerk W,Tayapiwatana C

更新日期：2003-12-01 00:00:00

abstract：:Using a codon-optimized gene fragment, we report remarkable yields for extracellular domain of human NK cell receptor (NKp30ex) when produced on M9 minimal medium, even with low (2g/L) glucose concentration. The yields were identical using media containing (15)NH(4)Cl or (15)NH(4)Cl in combination with all-(13)C-d-glu...

journal_title：Protein expression and purification

authors： Grave L,Tůmová L,Mrázek H,Kavan D,Chmelík J,Vaněk O,Novák P,Bezouška K

更新日期：2012-12-01 00:00:00

abstract：:Acyl-acyl carrier protein synthase (Aas) is widely used to synthesize thioester adducts of fatty acids between 8 and 18 carbons in length enzymatically to the phosphopantetheine group of acyl carrier protein. The enzyme is an 80.6-kDa inner membrane protein that functions in vivo as a 2-acylglycerophosphoethanolamine ...

journal_title：Protein expression and purification

authors： Shanklin J

更新日期：2000-04-01 00:00:00

abstract：:Ricin A chain (RTA) mutants which had been modified by the addition of three lysine residues, three lysines and an alanine, or six histidine residues at the carboxyl terminus were expressed in Escherichia coli. The recombinant proteins were purified to homogeneity by ion-exchange chromatography on CM-Sepharose CL-6B. ...

journal_title：Protein expression and purification

authors： Simpson JC,Roberts LM,Lord JM

更新日期：1995-10-01 00:00:00

abstract：:Expression of recombinant proteins in bacterial or eukaryotic systems often results in aggregation rendering them unavailable for biochemical or structural studies. Protein aggregation is a costly problem for biomedical research. It forces research laboratories and the biomedical industry to search for alternative, mo...

journal_title：Protein expression and purification

authors： Pullara F,Guerrero-Santoro J,Calero M,Zhang Q,Peng Y,Spåhr H,Kornberg GL,Cusimano A,Stevenson HP,Santamaria-Suarez H,Reynolds SL,Brown IS,Monga SP,Van Houten B,Rapić-Otrin V,Calero G,Levine AS

更新日期：2013-02-01 00:00:00

abstract：:Acetyl-CoA C-acetyltransferase synthase gene (AACT) cDNA, DNA and promoter were cloned from Sanghuangporus baumii. The gene ORF (1260 bp) encoded 419 amino acids. The AACT DNA includes five exons (1-84 bp, 140-513 bp, 570-1027 bp, 1090-1282 bp, 1344-1494 bp) and four introns (85-139 bp, 514-569 bp, 1028-1089 bp, 1283-...

journal_title：Protein expression and purification

authors： Wang X,Wang S,Xu X,Sun J,Ma Y,Liu Z,Sun T,Zou L

更新日期：2020-06-01 00:00:00

abstract：:Conkunitzin-S1 from the cone snail Conus striatus is the first member of a new neurotoxin family with a canonical Kunitz domain fold. Conk-S1 is 60 amino acids long and lacks one of the three conserved disulfide bonds typically found in Kunitz domain modules. It binds specifically to voltage activated potassium channe...

journal_title：Protein expression and purification

authors： Bayrhuber M,Graf R,Ferber M,Zweckstetter M,Imperial J,Garrett JE,Olivera BM,Terlau H,Becker S

更新日期：2006-06-01 00:00:00

abstract：:Folding and assembly studies with alpha-helical membrane proteins are often hampered by the absence of high-level expression systems as well as by missing suitable in vitro refolding procedures. Experimental constraints and requirements for heterologous expression and in vitro assembly of cytochrome b6 have been exami...

journal_title：Protein expression and purification

authors： Prodöhl A,Dreher C,Hielscher R,Hellwig P,Schneider D

更新日期：2007-12-01 00:00:00

abstract：:Human cystatin C (HCC), encoded by cystatin 3 gene, is a 13.3kDa endogenous cysteine proteinase inhibitor and an important biomarker of renal function. However, expressing recombinant cystatin C is difficult because of low yield and inclusion bodies in Escherichia coli (E. coli). In this study, we cloned HCC gene into...

journal_title：Protein expression and purification

authors： Zhou Y,Zhou Y,Li J,Chen J,Yao Y,Yu L,Peng D,Wang M,Su D,He Y,Gou L

更新日期：2015-07-01 00:00:00

abstract：:Der p 2, a major allergen derived from the house dust mite Dermatophagoides pteronyssinus, is one of the most clinically relevant allergens worldwide. Recombinant Der p 2 (rDer p 2) is useful in clinical diagnosis and disease-specific immunotherapy. However, previous studies showed that Der p 2 can only be expressed i...

journal_title：Protein expression and purification

authors： Cao T,Zhang Z,Liu ZG,Dou X,Zhang J,Zhang W,Wu B,Yu ZD,Wei Z,Yu B

更新日期：2016-05-01 00:00:00

abstract：:MMP1 is an essential enzyme for tissue remodeling both in normal and pathological states. We report a method of purifying activated human MMP1 in E. coli without using urea or 4-Aminophenylmercuric acetate (APMA). Instead, a non-ionic detergent, Triton X-100, was used in the lysis buffer to solubilize MMP1 followed by...

journal_title：Protein expression and purification

authors： Kumar L,Colomb W,Czerski J,Cox CR,Sarkar SK

更新日期：2018-08-01 00:00:00

abstract：:Recombinant murine MRP14 (mMRP14) was produced in Escherichia coli using the pGEX expression system. The mass of fusion protein, by electrospray ionization-mass spectrometry (ESI/MS), was 39,213 Da which compares well with the theoretical mass (39,210.4 Da). Thrombin digestion of fusion protein was expected at a clone...

journal_title：Protein expression and purification

authors： Raftery MJ,Collinson L,Geczy CL

更新日期：1999-03-01 00:00:00

abstract：:Amyloid-beta (Aβ) peptide mediates several neurodegenerative diseases. The 42 amino acid (Aβ1-42) is the predominant form of peptide found in the neuritic plaques and has been demonstrated to be neurotoxic in vivo and in vitro. The availability of large quantities of Aβ peptide will help in several biochemical and bio...

journal_title：Protein expression and purification

authors： Chhetri G,Pandey T,Chinta R,Kumar A,Tripathi T

更新日期：2015-10-01 00:00:00

abstract：:PLD's (Phospholipases D) are ubiquitously expressed proteins involved in many transphosphatidylation reactions. They have a bi-lobed structure composed by two similar domains which at their interface reconstitute the catalytic site through the association of the two conserved HxKx(4)Dx(6)GSxN motifs. PLD1 interacts wi...

journal_title：Protein expression and purification

authors： Viparelli F,Doti N,Sandomenico A,Marasco D,Dathan NA,Miele C,Beguinot F,Monti SM,Ruvo M

更新日期：2008-06-01 00:00:00

abstract：:L-Threonine dehydrogenase was purified 10,000-fold to a specific activity approximately 300 mumol.min-1.mg-1 protein from porcine liver mitochondria. Purification to apparent homogeneity was achieved by sequential chromatography on DEAE Sepharose FF, Affi-Gel Blue, Sephacryl S-200, Matrex Gel Red A, and Matrex Gel Gre...

journal_title：Protein expression and purification

authors： Kao YC,Davis L

更新日期：1994-10-01 00:00:00

abstract：:Adrenal cytochrome b561 (cyt b561) is the prototypical member of an emerging family of proteins that are distributed widely in vertebrate, invertebrate and plant tissues. The adrenal cytochrome is an integral membrane protein with two b-type hemes and six predicted transmembrane helices. Adrenal cyt b561 is involved i...

journal_title：Protein expression and purification

authors： Liu W,Rogge CE,Kamensky Y,Tsai AL,Kulmacz RJ

更新日期：2007-12-01 00:00:00

abstract：:Soluble guanylate cyclase (sGC) has been purified from 100 L cell culture infected by baculovirus using the newer and highly effective titerless infected-cells preservation and scale-up (TIPS) method. Successive passage of the enzyme through DEAE, Ni(2+)-NTA, and POROS Q columns obtained approximately 100mg of protein...

journal_title：Protein expression and purification

authors： Emmons TL,Mathis KJ,Shuck ME,Reitz BA,Curran DF,Walker MC,Leone JW,Day JE,Bienkowski MJ,Fischer HD,Tomasselli AG

更新日期：2009-06-01 00:00:00

abstract：:Enzymatic hydrolysis of the N-iminylamide was investigated in this study. An enzyme possessing N-iminylamidase activity from pig liver was purified to electrophoretic homogeneity. This enzyme was also active, however, with imides and appears to be identical to pig liver imidase. The identification was confirmed by cop...

journal_title：Protein expression and purification

authors： Huang CY,Yang YS

更新日期：2005-03-01 00:00:00

abstract：:In microorganisms and plants, mevalonate kinase is involved in the biosynthesis of isoprenoid derivatives, one of the largest groups of natural products. We subcloned the gene of mevalonate kinase from Methanococcus jannaschii into a bacterial expression vector pLM1 with six continuous histidine codons attached to the...

journal_title：Protein expression and purification

authors： Chu X,Liu X,Yau M,Leung YC,Li D

更新日期：2003-08-01 00:00:00

abstract：:We describe a single method to purify milligram amounts of authentic wild-type and mutant HIV-1 and HIV-2 Tat proteins overexpressed in Escherichia coli. This method takes advantage of the highly basic, positively charged RNA binding domain present in both HIV-1 and HIV-2 Tat, which also facilitated purification of HI...

journal_title：Protein expression and purification

authors： Orsini MJ,García-Martínez LF,Mavankal G,Gaynor RB,Debouck CM

更新日期：1996-09-01 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：:The purpose of this work was to characterize the functions of two sesquiterpene synthase genes from moso bamboo (Phyllostachys edulis). Two novel sesquiterpene synthase genes, belonging to the Tpsa subfamily, were isolated from moso bamboo. MoTPS2 was 1641 bp in length and encoded a protein of 63 kDa, whereas MoTPS6 w...

journal_title：Protein expression and purification

authors： Chen X,Wang Y,Sun J,Wang J,Xun H,Tang F

更新日期：2016-04-01 00:00:00

abstract：:α-N-Acetylgalactosaminidase (α-GalNAc-ase; EC.3.2.1.49) is an exoglycosidase specific for the hydrolysis of terminal α-linked N-acetylgalactosamine in various sugar chains. The cDNA corresponding to the α-GalNAc-ase gene was cloned from Aspergillus niger, sequenced, and expressed in the yeast Saccharomyces cerevisiae....

journal_title：Protein expression and purification

authors： Mrázek H,Benada O,Man P,Vaněk O,Křen V,Bezouška K,Weignerová L

更新日期：2012-01-01 00:00:00

abstract：:Insoluble expression of intrinsically soluble proteins with native activity is potentially a promising alternative to soluble expression of folded protein or insoluble expression of unfolded protein requiring refolding. Here, we attempted to express highly soluble halophilic His-rich metal binding protein (HP) as inso...

journal_title：Protein expression and purification

authors： Tokunaga M,Arakawa T,Tokunaga Y,Sugimoto Y,Ishibashi M

更新日期：2019-04-01 00:00:00

abstract：:The UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase from Escherichia coli, an enzyme involved in the biosynthesis of the bacterial peptidoglycan monomer unit, was overproduced and purified to homogeneity on a large scale, yielding 4 mg of protein per liter of bacterial culture. Crystals of the complex with the subst...

journal_title：Protein expression and purification

authors： Auger G,Martin L,Bertrand J,Ferrari P,Fanchon E,Vaganay S,Pétillot Y,van Heijenoort J,Blanot D,Dideberg O

更新日期：1998-06-01 00:00:00

abstract：:The aim of this study was to evaluate the parameters that affect the production of the recombinant 10 kDa culture filtrate protein (CFP10), a promising reagent of high specificity for intradermoreaction and other antigen-based methods used in the diagnosis of tuberculosis. Conditions of Escherichia coli growth tempera...

journal_title：Protein expression and purification

authors： Dela Coletta Troiano Araújo L,Wibrantz M,Rodríguez-Fernández DE,Karp SG,Talevi AC,Maltempi de Souza E,Soccol CR,Thomaz-Soccol V

更新日期：2019-02-01 00:00:00

abstract：:L1 is a human cell adhesion glycoprotein involved in the development of the central nervous system that comprises six immunoglobulin-like domains (Ig1-Ig6), five fibronectin-type III (FN1-FN5) domains, a single transmembrane region and a cytoplasmic domain. It contains 20 potential N-glycosylation sites and is heavily...

journal_title：Protein expression and purification

authors： Gouveia RM,Morais VA,Peixoto C,Sousa M,Regalla M,Alves PM,Costa J

更新日期：2007-03-01 00:00:00

abstract：:The kinetic locking-on strategy improves the selectivity of protein purification procedures based on immobilized cofactor derivatives through use of enzyme-specific substrate analogues in irrigants to promote biospecific adsorption. This paper describes the development and application of this strategy to the one-chrom...

journal_title：Protein expression and purification

authors： O'flaherty M,McMahon M,Mulcahy P

更新日期：1999-02-01 00:00:00

abstract：:Interferon α-2a (IFNA2) is a member of the Type I interferon cytokine family, known for its antiviral and anti-proliferative functions. The role of this family in the innate immune response makes it an attractive candidate for the treatment of many viral and chronic immune-compromised diseases. Recombinant IFNA2 is cl...

journal_title：Protein expression and purification

authors： Bis RL,Stauffer TM,Singh SM,Lavoie TB,Mallela KM

更新日期：2014-07-01 00:00:00