Large-scale expression and purification of a soluble form of the pleckstrin homology domain of the human protooncogenic serine/threonine protein kinase PKB (c-akt) in Escherichia coli.

abstract：:Peptide-based affinity tags are commonly used in recombinant production/purification of proteins, and are often preceded or followed by a protease recognition sequence to allow tag removal. We describe a rat monoclonal antibody 2H5 recognizing an undecapeptide tag called "eTev", which contains a recognition sequence f...

journal_title：Protein expression and purification

authors： Tabata S,Kitago Y,Fujii Y,Mihara E,Tamura-Kawakami K,Norioka N,Takahashi K,Kaneko MK,Kato Y,Takagi J

更新日期：2018-07-01 00:00:00

abstract：:A strategy for the purification and cleavage of chimeric recombinant proteins based on a genetically engineered metal-binding peptide and a human renin cleavage site is described. Vectors were constructed to direct the synthesis of chimeric human immunodeficiency virus (HIV) reverse transcriptase (RT) or beta-galactos...

journal_title：Protein expression and purification

authors： Evans DB,Tarpley WG,Sharma SK

更新日期：1991-04-01 00:00:00

abstract：:Most parasitic protozoa lack the de novo purine biosynthetic pathway and rely exclusively on the salvage pathway for their purine nucleotide requirements. Enzymes of the salvage pathway are, therefore, candidate drug targets. We have cloned the Plasmodium falciparum adenylosuccinate synthetase gene. In the parasite, a...

journal_title：Protein expression and purification

authors： Jayalakshmi R,Sumathy K,Balaram H

更新日期：2002-06-01 00:00:00

abstract：:Sphingomyelinase C (SMC) of the actinomycete, Streptomycesgriseocarneus NBRC13471, was constitutively expressed to high levels using Streptomyces lividans host and thereafter was extracellularly secreted into the cell culture. Purified SMC had a high specific activity (approximately 550-950 U/mg) and was obtained in h...

journal_title：Protein expression and purification

authors： Sugimori D,Matsumoto Y,Tomita Y,Murayama K,Ogino C

更新日期：2012-02-01 00:00:00

abstract：:Fusion proteins expressed in bacteria are often insoluble or inefficiently purified by standard procedures previously reported to work well for the non-fused proteins. We report here a simple but general procedure that can be used to quickly customize and optimize the purification of milligram quantities of most GST f...

journal_title：Protein expression and purification

authors： Mercado-Pimentel ME,Jordan NC,Aisemberg GO

更新日期：2002-11-01 00:00:00

abstract：:The guanine nucleotide exchange factor EF-1beta gene from the thermoacidophilic archaeon Sulfolobus solfataricus (SsEF-1beta) was amplified by PCR and cloned into the pT7-7 expression vector. One of four selected clones harbored the T160C nucleotide substitution leading to the Y54H amino acid change in a hydrophobic r...

journal_title：Protein expression and purification

authors： Ianniciello G,Masullo M,Raimo G,Arcari P,Bocchini V

更新日期：1998-02-01 00:00:00

abstract：:Targeted anti-tumor toxins consist of a toxic functional moiety that is chemically linked or recombinantly fused to a cell-directing ligand. Ribosome-inactivating proteins (RIPs), especially type I RIPs such as saporin or dianthin, are commonly used as toxin components. Although expression of type I RIP-based fusion p...

journal_title：Protein expression and purification

authors： Gilabert-Oriol R,Thakur M,Weise C,Dernedde J,von Mallinckrodt B,Fuchs H,Weng A

更新日期：2013-09-01 00:00:00

abstract：:The human melanocortin 4 receptor (MC4r) was successfully expressed in Sf9 cells using the baculovirus infection system. N- and C-terminally His-tagged receptors generated B(max) values of 14 and 23 pmol receptor/mg membrane protein, respectively. The highest expression level obtained with the C-terminally His-tagged ...

journal_title：Protein expression and purification

authors： Dolby V,Collén A,Lundqvist A,Cronet P

更新日期：2004-10-01 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：:A new method for the purification of manganese superoxide dismutase from human liver is described. The procedure involves essentially three steps: DEAE-cellulose, hydroxylapatite, and butyl-Toyopearl chromatographies. The method has several advantages: (i) its simplicity and rapidity (it takes less than 3 days), (ii) ...

journal_title：Protein expression and purification

authors： Matsuda Y,Suzuki K,Ookawara T,Nakata T,Seo HG,Kawata S,Tarui S,Deutsch HF,Taniguchi N

更新日期：1991-04-01 00:00:00

abstract：:Periplasmic expression of recombinant proteins ensures the production of biologically active proteins in a correctly folded state with several key advantages. This research focused on the in-frame cloning of rhIL-15 in pET-20 (+) vector with pelB-leader sequence to direct the protein to the bacterial periplasm. The ta...

journal_title：Protein expression and purification

authors： Ahmed N,Afroze B,Abbas R,Khan MA,Akram M,Tahir S,Bakht S,Munir A,Shahid AA

更新日期：2021-01-01 00:00:00

abstract：:Deoxynivalenol (DON), a mycotoxin produced by several Fusarium species, is a worldwide contaminant of food and feedstuffs. The DON-specific single-chain variable fragment (scFv) antibody was produced in recombinant Escherichia coli. The variable regions of the heavy chain (V(H)) and light chain (V(L)) cloned from the ...

journal_title：Protein expression and purification

authors： Choi GH,Lee DH,Min WK,Cho YJ,Kweon DH,Son DH,Park K,Seo JH

更新日期：2004-05-01 00:00:00

abstract：:Post-translational modification of proteins is a dynamic way of generating new protein-protein interaction interfaces that are critical for signaling networks in diverse cellular functions. Purified recombinant proteins frequently lack these signature modifications. Using the tumor suppressor p53 as the model protein,...

journal_title：Protein expression and purification

authors： Acharya A,Xu XJ,Husain-Ponnampalam RD,Hoffmann-Benning S,Kuo MH

更新日期：2005-06-01 00:00:00

abstract：:The TCL1 gene, which is located on chromosome 14, plays a major role in human hematopoietic malignancies and encodes a 14-kDa protein whose function has not been determined. This gene is expressed in pre-B cells, in immature thymocytes, and, at low levels, in activated T cells but not in peripheral mature B cells and ...

journal_title：Protein expression and purification

authors： Du Bois GC,Song SP,Kulikovskaya I,Rothstein JL,Germann MW,Croce CM

更新日期：2000-04-01 00:00:00

abstract：:The major capsid protein L1 of human papillomavirus (HPV) contains the immunodominant neutralization epitopes of the virus and can auto-assembles to form virus-like particles (VLPs). Therefore, HPV L1 capsid proteins have been well investigated as potential vaccine candidates. To express large quantities of human papi...

journal_title：Protein expression and purification

authors： Ma Z,Chen B,Zhang F,Yu M,Liu T,Liu L

更新日期：2007-11-01 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：:A soluble N-terminal domain of the Escherichia coli adenylyl transferase (ATase) is responsible for deadenylylation activity of the intact enzyme. Previous studies of the deadenylylation activity have involved a fragment, AT-N423 (residues 1 to 423), which was extended by 17 amino acids to give AT-N440. This new domai...

journal_title：Protein expression and purification

authors： Xu Y,Wen D,Clancy P,Carr PD,Ollis DL,Vasudevan SG

更新日期：2004-03-01 00:00:00

abstract：:Magnolia officinalis Rehder et Wilson is a traditional Chinese herbal medicine that is used to treat various diseases such as neurosis, anxiety, and stroke. The main secondary metabolites in magnolia bark are phenolic compounds and terpenoids. Squalene synthase plays a significant role in catalyzing two farnesyl dipho...

journal_title：Protein expression and purification

authors： Zha L,Liu S,Su P,Yuan Y,Huang L

更新日期：2016-04-01 00:00:00

abstract：:To facilitate studies of multicomponent protein complexes, I have developed an Escherichia coli expression system which coexpresses up to four polypeptides from a single plasmid. The modular nature of the system enables efficient subcloning of a gene into each of the 4 cassettes in the polycistronic expression vector....

journal_title：Protein expression and purification

authors： Tan S

更新日期：2001-02-01 00:00:00

abstract：:Structural biology places a high demand on proteins both in terms of quality and quantity. Although many protein expression and purification systems have been developed, an efficient and simple system which can be easily adapted is desirable. Here, we report a new system which combines improved expression, solubility ...

journal_title：Protein expression and purification

authors： Liu H,Naismith JH

更新日期：2009-02-01 00:00:00

abstract：:PDZ (PSD-95, DiscsLarge, ZO1) domains function in nature as protein binding domains within scaffold and membrane-associated proteins. They comprise ∼90 residues and make specific, high affinity interactions with complementary C-terminal peptide sequences, with other PDZ domains, and with phospholipids. We hypothesized...

journal_title：Protein expression and purification

authors： Walkup WG 4th,Kennedy MB

更新日期：2014-06-01 00:00:00

abstract：:An Escherichia coli expression system that exploits the bacterial alkaline phosphatase (PhoA) signal sequence to translocate recombinant human epidermal growth factor (hEGF) to the periplasm was used to evaluate how changes in the composition and sequence of amino acids near the PhoA-hEGF junction influence the peripl...

journal_title：Protein expression and purification

authors： Campion SR,Elsasser E,Chung R

更新日期：1997-08-01 00:00:00

abstract：:In order to obtain bioactive α-bungarotoxin (αBtx) using recombinant protein technique, a codon-optimized synthetic gene was expressed in fusion with the N-terminal 10-His-tag and C-terminal Strep-tag in Escherichia coli. Further optimization through site-directed mutagenesis enabled moderate expression of the protein...

journal_title：Protein expression and purification

authors： Xu J,Li J,Wu X,Song C,Lin Y,Shen Y,Ye W,Sun C,Wang X,Li Z,Liu Y,Wei L,Li Z,Xu Z

更新日期：2015-06-01 00:00:00

abstract：:Darbepoetin alfa is an engineered and hyperglycosylated analog of recombinant human erythropoietin (EPO) which is used as a drug in treating anemia in patients with chronic kidney failure and cancer. This study desribes the secretory expression of a codon-optimized recombinant form of darbepoetin alfa in Leishmania ta...

journal_title：Protein expression and purification

authors： Kianmehr A,Golavar R,Rouintan M,Mahrooz A,Fard-Esfahani P,Oladnabi M,Khajeniazi S,Mostafavi SS,Omidinia E

更新日期：2016-02-01 00:00:00

abstract：:Ndt80 is a Saccharomyces cerevisiae meiosis-specific transcription factor responsible for promoting the stage-specific expression of a family of genes referred to as middle sporulation genes. Many members of this gene family are essential for the completion of meiotic chromosome segregation. Thus, Ndt80 is essential f...

journal_title：Protein expression and purification

authors： Sopko R,Stuart DT

更新日期：2004-01-01 00:00:00

abstract：:Peptide deformylases (PDFs) catalyze the removal of the formyl group from the N-terminal methionine residue in nascent polypeptide chains in prokaryotes. Its deformylation activity makes PDF an attractive candidate for the biocatalytic deprotection of formylated peptides that are used in chemoenzymatic peptide synthes...

journal_title：Protein expression and purification

authors： Di Toma C,Sonke T,Quaedflieg PJ,Volker Wagner AF,Janssen DB

更新日期：2013-05-01 00:00:00

abstract：:In Pisum sativum, the short-chain alcohol dehydrogenase-like protein (SAD) gene family consists of at least three members (SAD-A, -B, and -C). Expression of two of these genes (SAD-A and -C) in Escherichia coli or Pichia pastoris resulted in full-length soluble proteins. Purified SAD-A was used as antigen for antibody...

journal_title：Protein expression and purification

authors： Scherbak N,Brosché M,Ala-Häivälä A,Strid H,Ohrfelt A,Nilsson F,Strid A

更新日期：2009-01-01 00:00:00

abstract：:Transcription regulation in the cell occurs in the context of chromatin. It follows that a thorough investigation of the mechanism of transcription regulation must take into account the role of chromatin structure. Through classical and molecular genetic experiments in yeast, great strides have been made in understand...

journal_title：Protein expression and purification

authors： Pilon J,Terrell A,Laybourn PJ

更新日期：1997-06-01 00:00:00

abstract：:Organophosphorus hydrolase (OPH) is a ∼38kDa enzyme encoded by opd gene of Flavobacterium sp. The enzyme can hydrolyze and inactivate variety of organophosphate (OP)-compounds, including chemical warfare nerve agents. Thus, OPH is a strong candidate for the development of therapeutic intervention against OP-poisoning ...

journal_title：Protein expression and purification

authors： Satvik Iyengar AR,Tripathy RK,Bajaj P,Pande AH

更新日期：2015-07-01 00:00:00

abstract：:Human Fas ligand is a medically important membrane glycoprotein that induces the apoptosis of harmful cells. A new secretory expression and purification method was devised for the production of a large amount of recombinant human Fas ligand extracellular domain (hFasLECD) by Pichia pastoris. The expression plasmid con...

journal_title：Protein expression and purification

authors： Muraki M

更新日期：2006-12-01 00:00:00