Abstract:
:The whole encoding sequence for Yersinia pestis LcrV antigen was cloned into pET-32a(+) and expressed in Escherichia coli BL21 (DE3). The LcrV was high level expressed in the E. coli cytoplasm in a completely soluble form. Recombinant LcrV could be purified from the supernatant of the bacteria lysate after chromatography using a combination of Phenyl-Sepharose F F, DEAE-Sepharose F F and Hiload Superdex 75. The final yield of approximately 3 g of purified rLcrV from 42 L bioreactor containing 25 L LB medium was obtained. High-titer IgG directed against rLcrV was detected positive after immunization on the BALB/c mice. The results presented here exhibit the ability to generate multi-gram quantities of non-tagged rLcrV from E. coli that can be used for the development of vaccine for preventing plague.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
li J,Li B,Li G,Ren J,Zhang J,Xu C,Yang X,Liu S,Fu L,Chen Wdoi
10.1016/j.pep.2007.10.011subject
Has Abstractpub_date
2008-02-01 00:00:00pages
136-42issue
2eissn
1046-5928issn
1096-0279journal_volume
57pub_type
杂志文章abstract::Granulins (GRNs) are potent growth factors that are upregulated in many aggressive cancers from a wide range of organs. GRNs form tight, disulphide bonded, beta hairpin stacks, making them difficult to express in recombinant form. We recently described Ov-GRN-1, a GRN family member secreted by the carcinogenic liver f...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.06.018
更新日期:2011-10-01 00:00:00
abstract::The aim of this study was to establish a method of purifying intact complement factor H (CFH) from human plasma. CFH was isolated from human plasma by polyethylene glycol (PEG) precipitation, following three sequential chromatographic columns, which consisted of l-lysine Sepharose column, Resource Q column and Sephacr...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2013.07.014
更新日期:2013-10-01 00:00:00
abstract::Silent information regulator 2 (Sir2) proteins are a class of protein deacetylase enzymes that play key roles in transcriptional gene silencing, DNA repair, and aging. Here, we describe the high-level bacterial expression and purification of a human SirT2 construct that yields high resolution NMR spectra. By removing ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.12.006
更新日期:2006-07-01 00:00:00
abstract::Glycine N-acyltransferase (GLYAT) is a phase II metabolic detoxification enzyme for exogenous (xenobiotic) and endogenous carboxylic acids; consisting of fatty acids, benzoic acid, and salicylic acid. GLYAT catalyzes the formation of hippurate (N-benzoylglycine) from the corresponding glycine and benzoyl-CoA. Herein, ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.02.007
更新日期:2014-05-01 00:00:00
abstract::Transmembrane and coiled-coil domains 1 (TMCO1) has a highly conserved amino acid sequence among species, indicating a critical role of TMCO1 in cell physiology. The deficiency of TMCO1 in humans is associated with cerebrofaciothoracic dysplasia (CFTD), glaucoma, osteogenesis and the occurrence of cancer. TMCO1 was re...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105803
更新日期:2021-03-01 00:00:00
abstract::Escherichia coli glutamic acid decarboxylase is a pyridoxal phosphate-dependent enzyme that catalyzes the alpha-decarboxylation of glutamate to yield 4-amino-butyrate and CO2. The E. coli chromosome contains two genes encoding for this enzyme, gadA and gadB, which map at distinct loci. Their protein products differ in...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0121
更新日期:1996-12-01 00:00:00
abstract::Hereditary enzymopenic methemoglobinemia is a rare disease that predominantly results from defects in either the erythrocytic (type I) or microsomal (type II) forms of the enzyme NADH:cytochrome b5 reductase (EC 1.6.2.2). All 25 currently identified type I and type II methemoglobinemia mutants have been expressed in E...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00046-9
更新日期:2003-07-01 00:00:00
abstract::This paper reports the overproduction and the details of a rapid method to purify active sigmaS monomers from a T7 RNA polymerase-based protein expression system. This 2-day procedure involves solubilizing inclusion bodies in sarkosyl detergent, removal of sarkosyl by dialysis, and a single gel filtration column chrom...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0069
更新日期:1996-08-01 00:00:00
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journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1994.1014
更新日期:1994-02-01 00:00:00
abstract::The leucine aminopeptidase from Aeromonas proteolytica (also known as Vibrio proteolyticus) (AAP) is a metalloenzyme with broad substrate specificity. The open reading frame (ORF) for AAP encodes a 54 kDa enzyme, however, the extracellular enzyme has a molecular weight of 43 kDa. This form of AAP is further processed ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.05.004
更新日期:2004-10-01 00:00:00
abstract::Aquaporins are integral membrane channel proteins found in all kingdoms of life. The Escherichia coli aquaporin Z (AqpZ) has been shown to solely conduct water at high permeability. Functional AqpZ is generally purified from the membrane fraction. However, the quantity of the purified protein is limited. In this study...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.08.014
更新日期:2015-11-01 00:00:00
abstract::The protooncogenic serine/threonine protein kinase PKB contains an amino-terminal pleckstrin homology (PH) domain which binds phosphatidylinositides. The PH domain, composed of approximately 100 loosely conserved amino acids, is found in many proteins, including kinases, phospholipases C, GTPases, GTPase-activating pr...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1999.1120
更新日期:1999-11-01 00:00:00
abstract::Bovine follicle-stimulating hormone (bFSH) is a pituitary gonadotropin composed of two non-covalently associated polypeptide subunits, which must be glycosylated, folded, and assembled as a heterodimer to be biologically active. Low-level expression of the recombinant bFSH is the factor that limits its usefulness as a...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.03.016
更新日期:2007-08-01 00:00:00
abstract::To expand the application of the streptavidin-biotin technology for reversible affinity purification of biotinylated proteins, a novel form of monomeric streptavidin was engineered and produced using Bacillus subtilis as the expression host. By changing as little as two amino acid residues (T90 and D128) to alanine, t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00021-9
更新日期:2002-08-01 00:00:00
abstract::Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit epsilon N-methyltransferase (Protein methylase III, Rubisco LSMT, EC 2.1.1.43) catalyzes methylation of the epsilon-amino group of Lys-14 in the large subunit of Rubisco. In this paper, an affinity purification procedure for pea (Pisum sativum L. cv Laxton'...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1995.1070
更新日期:1995-08-01 00:00:00
abstract::To understand the key processes of cell-free protein synthesis, the synthesis of adipose-type fatty acid binding protein (A-FABP) by a rapid translation system was examined under various conditions. The synthesis of A-FABP was achieved by using an expression vector of A-FABP containing a T7 promoter. However, synthesi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1570
更新日期:2002-03-01 00:00:00
abstract::The zoonotic Rift Valley fever virus affects livestock and humans in Africa and on the Arabian Peninsula. The economic impact of this pathogen due to livestock losses, as well as its relevance to public health, underscores the importance of developing effective and easily distributed vaccines. Vaccines that can be del...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2016.07.003
更新日期:2016-11-01 00:00:00
abstract::Novel GSH-AP (phenoxyl agarose coated gigaporous polystyrene, Agap-co-PSt) microspheres were successfully prepared by introducing GSH ligand into hydrophilic AP microspheres pre-activated with 1,4-butanediol diglycidyl ether. The gigaporous structure and chromatographic properties of GSH-AP medium were evaluated and c...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2013.11.004
更新日期:2014-03-01 00:00:00
abstract::The high specific lysyl endopeptidase (Lys-C; EC 3.4.21.50) is often used for the initial fragmentation of polypeptide chains during protein sequence analysis. However, due to its specificity it could be a useful tool for the production of tailor-made protein hydrolysates with for example bioactive or techno functiona...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.09.024
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abstract::An improved, simplified method for the purification of recombinant, tagged DnaA proteins is described. The presented protocol allowed us to purify soluble DnaA proteins from two different bacterial species: Helicobacter pylori and Streptomyces coelicolor, but it can most likely also be used for the isolation of DnaA p...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.01.010
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abstract::We have cloned, from total human liver RNA, the cDNA encoding apolipoprotein E3 (apoE3). Site-directed mutagenesis was used to obtain the cDNA encoding the apoE4 isoform, a major variant of this apolipoprotein in man. These two cDNAs were subcloned into the procaryotic expression vector pAHRS. A polyhistidine tag was ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0061
更新日期:1996-06-01 00:00:00
abstract::The bivalent anti-human anti-T cell immunotoxin A-dmDT390-bisFv(G(4)S) was developed for treatment of T cell leukemia, autoimmune diseases, and tolerance induction for transplantation. The multi-domain structure of the bivalent immunotoxin hinders efficient production in Escherichia coli and most eukaryotes are sensit...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00009-8
更新日期:2002-07-01 00:00:00
abstract::An antifungal protein with a chitinase-like N-terminal sequence, designated delandin, was isolated from the rice bean. The protein exhibited a molecular weight of 28 kDa and was adsorbed on both blue Affi-Gel and SP-Toyopearl. It exerted antifungal action toward Mycosphaerella arachidicola, Botrytis cinerea, Fu- sariu...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1596
更新日期:2002-04-01 00:00:00
abstract::We have developed a family of cloning vectors that direct expression of fusion proteins that mimic aggregated immunoglobulin (IgG) (AIG) and immune complex function with respect to their interactions with FcgammaR and that allow for the inclusion and targeting of a second protein domain to cells expressing FcgammaR. T...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1406
更新日期:2001-04-01 00:00:00
abstract::Sarcotoxin IA is an antibacterial peptide that is secreted by a meat-fly Sarcophaga peregrina larva in response to a hypodermic injury or bacterial infection. This peptide is highly toxic against a broad spectrum of both Gram-positive and Gram-negative bacteria and lethal to microbes even at nanomolar concentrations. ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00697-6
更新日期:2003-04-01 00:00:00
abstract::Replacing the chymotrypsin inhibitory loop of soybean Bowman-Birk inhibitor (sBBI) with a VEGF binding peptide (BBI-AV) significantly reduces the overall purification yield when BBI-AV is produced as a fusion protein in a Bacillussubtilis expression system. The low purification yield is primarily due to a higher fract...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.08.006
更新日期:2009-12-01 00:00:00
abstract::An alpha-amylase has been purified from the thermophilic fungus Scytalidium thermophilum. A ninefold purification was achieved in a single step using fluidized bed chromatography wherein alginate was used as the affinity matrix. There are at least two isoenzymes as shown by concanavalin A (Con A)-agarose column chroma...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2000.1308
更新日期:2000-11-01 00:00:00
abstract::Manganese superoxide dismutase (Mn-SOD) is one of the major enzymes responsible for the defense against oxidative damage due to reactive oxygen species (ROS) in the mitochondria. The present study aimed to produce and evaluate the genetically engineered manganese superoxide dismutase protein. A recombinant plasmid con...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.12.008
更新日期:2011-05-01 00:00:00
abstract::Myrosinases are thioglucosidases that hydrolyze the natural plant products glucosinolates. We have expressed the myrosinase MYR1 from Brassica napus in Saccharomyces cerevisiae. The recombinant myrosinase was enzymatically active which shows that the MYR1, which in the plant is complex bound with myrosinase-binding pr...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1999.1158
更新日期:1999-12-01 00:00:00
abstract::Introduction and expression of foreign genes in bacteria often results accumulation of the foreign protein(s) in inclusion bodies (IBs). The subsequent processes of refolding are slow, difficult and often fail to yield significant amounts of folded protein. RHG1 encoded by rhg1 was a soybean (Glycine max L. Merr.) tra...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.12.017
更新日期:2007-06-01 00:00:00