Abstract：
:Rat granulocyte macrophage colony-stimulating factor (rGM-CSF) cDNA was amplified and cloned, and recombinant-rGM-CSF (R-rGM-CSF) was expressed and isolated from Escherichia coli. The synthesis of R-rGM-CSF was directed by a modified, inducible maltose binding protein (MBP) gene fusion expression vector, pMTR-23, and secreted to the periplasm. The vector pMTR-23 contains a new multiple cloning site encoding a unique thrombin-sensitive cleavage site and short spacer arm which facilitates separation of the MBP from the foreign protein domain of hybrid proteins. Biologically active R-rGM-CSF was rapidly purified by a combination of affinity and ion exchange chromatography, with a yield of 1.5 mg of R-rGM-CSF per liter of cultured cells. The purified R-rGM-CSF, like the native molecule, exhibits potent biological activity in two rGM-CSF-specific assays, considerably enhancing the accessory activity of rat dendritic cells and stimulating the differentiation of dendritic cells from fresh cultures of rat bone marrow cells. Although dendritic cells are difficult to isolate from tissues, the availability of R-rGM-CSF should now facilitate the development of large numbers of dendritic cells and the understanding of their regulatory role in the immune response.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Holowachuk EW,Ruhoff MSdoi
10.1006/prep.1995.1077subject
Has Abstractpub_date
1995-10-01 00:00:00pages
588-96issue
5eissn
1046-5928issn
1096-0279pii
S1046-5928(85)71077-7journal_volume
6pub_type
杂志文章abstract：:To produce a monoclonal antibody specific to a mouse major histocompatibility complex (MHC) class II protein, we synthesized the complementary DNAs for the heavy and light chains of a monoclonal antibody by PCR amplification. These cDNAs were then introduced separately into tobacco plant cells. After performing Northe...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.02.015
更新日期：2007-07-01 00:00:00
abstract：:Kynurenine 3-monooxygenase (KMO) is an enzyme central to the kynurenine pathway of tryptophan metabolism. KMO has been implicated as a therapeutic target in several disease states, including Huntington's disease. Recombinant human KMO protein production is challenging due to the presence of transmembrane domains, whic...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2013.11.015
更新日期：2014-03-01 00:00:00
abstract：:Developmentally regulated G-proteins (DRGs) are a highly conserved family of GTP-binding proteins found in archaea, plants, fungi and animals, indicating important roles in fundamental pathways. Their function is poorly understood, but they have been implicated in cell division, proliferation, and growth, as well as s...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.05.009
更新日期：2009-10-01 00:00:00
abstract：:A method for the simplified, reproducible production of both normal and altered versions of human proinsulin has been developed. A polyhistidine/proinsulin fusion protein was expressed using a prokaryotic expression system and partially purified by affinity chromatography. Disulfide bonds within the polypeptide were f...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1998.1024
更新日期：1999-04-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2009.03.010
更新日期：2009-08-01 00:00:00
abstract：:Human dermcidin, an anionic antimicrobial peptide expressed in the pons of the brain and the sweat glands, displays antimicrobial activity against pathogenic microorganisms such as Staphylococcus aureus and Candida albicans. Here, we describe the recombinant production of a 48 amino acid dermcidin variant with C-termi...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.07.002
更新日期：2006-02-01 00:00:00
abstract：:The second enzyme of the naphthalene degradation pathway in Pseudomonas putida G7 is NahB, a dehydrogenase that converts cis-1,2-dihydroxy-1,2-dihydronaphthalene to 1,2-dihydroxynaphthalene. We report the cloning, optimization of expression, purification, kinetic studies and preliminary structural characterization of ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2017.01.005
更新日期：2017-04-01 00:00:00
abstract：:17β-hydroxysteroid dehydrogenase type 1 (17β-HSD1) mainly catalyzes the reduction of estrone into estradiol. The enzymatic conversion is a critical step in estradiol accumulation in breast tissue, which is a valuable prognosis index of breast cancer disease. However, the source of 17β-HSD1 for inhibitor design is limi...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2017.06.015
更新日期：2017-09-01 00:00:00
abstract：:Mammalian target of rapamycin (TOR) controls cell growth and metabolism in response to the availability of nutrients, growth factors, and the cellular energy status. Misregulation of TOR can result in a pathogenic increase or decrease in organ size and in cancer. TOR can be inhibited by binding of a complex of rapamyc...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.01.014
更新日期：2008-05-01 00:00:00
abstract：:Staphylococcal enterotoxins (SEs) are powerful superantigens that stimulate non-specific T-cell proliferation produced by Staphylococcus aureus and draw considerable attention as ideal drugs for cancer therapy. The filtrate of S. aureus culture has been used as ampul named Staphylococcal enterotoxin C injection in cli...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.08.005
更新日期：2007-12-01 00:00:00
abstract：:The gene encoding a thermostable beta-glucosidase (cel3a) was isolated from the thermophilic fungus Talalaromyces emersonii by degenerate PCR and expressed in the filamentous fungus Trichoderma reesei. The cel3a gene encodes an 857 amino acid long protein with a calculated molecular weight of 90.59 kDa. Tal. emersonii...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.08.006
更新日期：2004-12-01 00:00:00
abstract：:The hepatitis E virus (HEV) capsid antigen has been proposed as a candidate subunit vaccine for the prevention of hepatitis E. The full-length HEV ORF2 protein product is predicted to contain 660 amino acids and to weigh 72,000 daltons. Expression of the HEV ORF2 capsid gene from recombinant baculoviruses in insect ce...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1997.0817
更新日期：1998-02-01 00:00:00
abstract：:Human HEK293 cells that stably express the Epstein Barr nuclear antigen 1 (EBNA1) support the episomal replication of plasmids containing the Epstein Barr virus origin of replication (EBV oriP). A 293EBNA (293E) cell line expressing the human corticotropin-releasing hormone receptor subtype I (CRHR1) from an episomal ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1996.0701
更新日期：1997-04-01 00:00:00
abstract：:During the screening for tyrosine phenol-lyase-producing thermophiles, we isolated an obligatory symbiotic thermophile, Symbiobacterium sp. SC-1, which grew only in coculture with Bacillus sp. SK-1. A gene encoding thermostable tyrosine phenol-lyase (TPL) was cloned from the genomic DNA of the Symbiobacterium sp. SC-1...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1997.0792
更新日期：1997-12-01 00:00:00
abstract：:Production of antibody fragments in heterologous hosts such as Escherichiacoli provides a unique and cost-effective method to develop engineered vectors for tumor targeting. A single-chain Fragment variable (scFv) of the murine monoclonal antibody MAb-B43.13 targeting CA125 in epithelial ovarian cancer was previously ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.07.007
更新日期：2014-10-01 00:00:00
abstract：:Refolding of proteins from inclusion bodies is affected by several factors, including solubilization of inclusion bodies by denaturants, removal of the denaturant, and assistance of refolding by small molecule additives. We will review key parameters associated with (1) conformation of the protein solubilized from inc...
journal_title：Protein expression and purification
pub_type： 杂志文章,评审
doi：10.1016/s1046-5928(02)00641-1
更新日期：2003-03-01 00:00:00
abstract：:Recombinant OSU VP4 protein, and outer capsid antigen of porcine rotavirus, was purified to a high level from the spent broth of baculovirus-infected Spodoptera frugiperda insect cells. Initial clarification of the broth with a 0-60% ammonium sulfate cut retained 93% of the total VP4. Q-sepharose ion exchange chromato...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1993.1030
更新日期：1993-06-01 00:00:00
abstract：:Borneol is a bicyclic plant monoterpene. It can be degraded by soil microorganisms through the conversion of borneol dehydrogenase (BDH) and a known camphor degradation pathway. Recombinant BDH from Pseudomonas sp. TCU-HL1 was produced in the form of inclusion body. The refolded BDH1 tends to precipitate. Insoluble re...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2020.105715
更新日期：2020-11-01 00:00:00
abstract：:Flt3 ligand (FL) is a potent hematopoietic cytokine that affects the growth and differentiation of hematopoietic progenitor and stem cells both in vivo and in vitro. Pichia pastoris transformants secreting high-level rhFL were obtained using 'yeastern blotting' method and the expression level in liquid was about 30 mg...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.04.001
更新日期：2005-08-01 00:00:00
abstract：:Rotavirus VP8* subunit is the minor trypsin cleavage product of the spike protein VP4, which is the major determinant of the viral infectivity and neutralization. To study the structure-function relationship of this fragment and to obtain type-specific reagents, substantial amounts of this protein are needed. Thus, fu...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.09.014
更新日期：2006-04-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2008.09.008
更新日期：2009-02-01 00:00:00
abstract：:Glutathione S-transferase (GST) fusion proteins are widely used in protein production for pure immunogens, protein-protein, and DNA-protein interaction studies. Using basic pGEX vectors, foreign DNA is introduced to the C-terminus of the GST gene and the produced fusion proteins are C-terminally orientated. However, b...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.11.012
更新日期：2005-04-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2015.12.008
更新日期：2016-04-01 00:00:00
abstract：:Accurate diagnosis is essential for the treatment, prevention, and control of tuberculosis. Poor specificity of the tuberculin skin test in BCG-vaccinated populations and constraints to implementation of PCR and CMI-based diagnostic assays in developing countries warrant development of easy-to perform robust serologic...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.04.014
更新日期：2005-11-01 00:00:00
abstract：:Cardiomyopathy-related mutations in human cardiac troponin subunits, including troponin C (hcTnC), troponin I (hcTnI), and troponin T (hcTnT), are well-documented. Recently, it has been recognised that human cardiac troponin (hcTn) is a sophisticated allosteric system. Therefore, the effect of drugs on this protein co...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.10.005
更新日期：2013-02-01 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：:A rabbit liver cDNA library in phage lambdagt10 was screened using the coding cDNA for human cytosolic serine hydroxymethyltransferase. A clone of 1754 bp was isolated and the nucleotide sequence showed an open reading frame of 1455 bp, which coded for rabbit cytosolic serine hydroxymethyltransferase and was flanked b...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1998.0890
更新日期：1998-07-01 00:00:00
abstract：:BetaB2-crystallin, the major subunit of beta-crystallins, is difficult to purify either from lens homogenate or from betaH-or betaL-crystallins. It has been prepared by heterologous expression in Escherichia coli. Most often, the methods used for purifying a recombinant globular protein employ the combination of ion-e...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00675-7
更新日期：2003-03-01 00:00:00
abstract：:Expression and purification of human beta-secretase (BACE1) in bacteria have been plagued with issues concerning solubility, inhomogeneous N-terminus, and lack of enzymic activity. Several forms of the mature human BACE1 have been expressed in Escherichia coli with different N-terminal extensions and without the C-ter...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2003.12.014
更新日期：2004-04-01 00:00:00
abstract：:The AP-1 transcription factor is a dimeric protein complex formed primarily between Jun (c-Jun, JunB, JunD) and Fos (c-Fos, FosB, Fra-1, Fra-2) family members. These distinct AP-1 complexes are expressed in many cell types and modulate target gene expression implicated in cell proliferation, differentiation, and stres...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.01.016
更新日期：2008-05-01 00:00:00
