Abstract：
:Recombinant baculoviruses have proved to be a very useful means to express many proteins over the last 20 years. Since their introduction, there have been a number of significant improvements that have simplified and speeded up the construction of baculoviruses. One of the most commonly used methods relies upon recombination with the baculovirus genome maintained in Escherichia coli. In this paper, we report the conversion of nearly all the steps in this process including the expression testing and purification to a multi-well plate format. This enables a significant increase in the number of constructs that can be processed in a shorter period of time and an order of magnitude increase in the number of expression conditions that can be analysed. A key step in our process is that the transfection is done in suspension rather than adherent cells, which gives a much higher virus titre than in the standard methods.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
McCall EJ,Danielsson A,Hardern IM,Dartsch C,Hicks R,Wahlberg JM,Abbott WMdoi
10.1016/j.pep.2005.03.021keywords:
subject
Has Abstractpub_date
2005-07-01 00:00:00pages
29-36issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(05)00109-9journal_volume
42pub_type
杂志文章abstract：:Antibacterial peptides from various sources express different antibacterial activity. In order to obtain a high activity antibacterial peptide, the sequences of four antimicrobial peptides--Protegrin-1, 4 kDa Scorpion Defensin, Metalnikowin-2A and Sheep Myeloid Antibacterial Peptide SMAP-29--were exploited to generate...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.08.006
更新日期：2008-01-01 00:00:00
abstract：:Thymosin α1-thymopentin (Tα1-TP5) fusion peptide has been proved to be an immune regulator based on its higher immunoregulatory activity than Tα1 and TP5. To obtain Tα1-TP5 more effectively and economically, Tα1-TP5 was genetically fused to a self-cleaving intein-chitin binding domain tag for purification via chitin b...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.04.013
更新日期：2012-07-01 00:00:00
abstract：:Hsp47 is regarded as a collagen-specific chaperone with several suggested roles in collagen biosynthesis under normal and disease conditions. We describe here a procedure for the expression and purification of Hsp47 in Escherichia coli using the IMPACT expression system (New England Biolabs) where the guest gene is fu...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2001.1470
更新日期：2001-10-01 00:00:00
abstract：:Soluble oligomers and fibrillar deposits of amyloid beta (Abeta) are key agents of Alzheimer's disease pathogenesis. However, the mechanism of amyloid aggregation and its interaction with live cells still remain unclear requiring the preparation of large amounts of pure and different Abeta peptides. Here we describe a...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.02.009
更新日期：2009-07-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2018.04.001
更新日期：2018-08-01 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
pub_type： 杂志文章
doi：10.1006/prep.1997.0741
更新日期：1997-08-01 00:00:00
abstract：:The assembly pathway of the insect cell Spodoptera frugiperda (Sf-9) was engineered to include expression of the murine chaperone immunoglobulin heavy chain binding protein (BiP) using the baculovirus vector. The impact of BiP coexpression on the production and secretion of functional and soluble recombinant immunoglo...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1994.1082
更新日期：1994-12-01 00:00:00
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 ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1995.1077
更新日期：1995-10-01 00:00:00
abstract：:A stable mammalian cell line expressing highly active bovine interferon-gamma (BoIFN-γ) was generated using Flp recombinase-mediated integration. This recombinant 293 cell line (B1) efficiently secreted FLAG-tagged BoIFN-γ protein into the culture supernatant, as determined by ELISA and Western blot. The recombinant B...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.04.012
更新日期：2014-07-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2019.01.001
更新日期：2019-04-01 00:00:00
abstract：:A growing number of studies require the purification of multiple proteins simultaneously and the development of simple economical high-throughput purification methods is essential. We have tested the purification of two related proteins in a variety of conditions to benchmark the semi-automated affinity chromatography...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.01.014
更新日期：2014-04-01 00:00:00
abstract：:An unknown d-tagatose 3-epimerase (DTE) containing a IoIE domain was identified and cloned from Escherichia coli. This gene was subcloned into the prokaryotic expression vector pET-15b, and induced by IPTG in E. coli BL21 expression system. Through His-select gel column purification and fast-protein liquid chromatogra...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.06.015
更新日期：2015-10-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2015.01.012
更新日期：2015-07-01 00:00:00
abstract：:Pen b 26 is one of the allergens produced by Penicillium brevicompactum which is one of the most prevalent in door airborne fungi and a major source of respiratory problems, including asthma. Pen b 26 wa scloned and expressed as an N-terminal as well as a C-terminal His6 tagged fusion protein in Escherichia coli. This...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.01.004
更新日期：2009-05-01 00:00:00
abstract：:Mammalian α-defensins contribute to innate immunity by exerting antimicrobial activity against various pathogens. To perform structural and functional analysis of α-defensins, large amounts of α-defensins are essential. Although many expression systems for the production of recombinant α-defensins have been developed,...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.04.007
更新日期：2015-08-01 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：: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
pub_type： 杂志文章
doi：10.1016/j.pep.2006.10.008
更新日期：2007-03-01 00:00:00
abstract：:Androgen receptor (AR)-associated coregulator 70 (ARA70) is a cytoplasmic protein that has been characterized to have the ability to induce AR transcriptional activity in response to androgens and anti-androgens in prostate cancer cells. AR has been shown to have an important role in the progression of prostate cancer...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.11.002
更新日期：2005-02-01 00:00:00
abstract：:The transcription factor IIIC (TFIIIC) is a multisubunit DNA-binding factor required for promoter recognition and TFIIIB assembly on tRNA genes transcribed by RNA polymerase III. Yeast TFIIIC consists of six subunits, organized in the two globular subcomplexes tauA and tauB, which recognize two internal tDNA promoter ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.06.013
更新日期：2006-02-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2003.11.001
更新日期：2004-03-01 00:00:00
abstract：:A simple and efficient method for hyperexpression in Escherichia coli and purification of capsid protein, p24, of human immunodeficiency virus type 1 (HIV-1) of both B- and C-subtypes is described. DNA-encoding p24 of C-subtype was cloned from C-subtype gag sequence which was obtained by PCR amplification using DNA ex...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1266
更新日期：2000-08-01 00:00:00
abstract：:The coding region for arginyl-tRNA synthetase from jack bean (Canavalia ensiformis) has been sequenced and cloned into the bacterial expression vector pET32a. Transformation of BL21 cells and induction with IPTG results in the high level expression of the protein fused N-terminally with thioredoxin and bearing a His-t...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.05.008
更新日期：2008-10-01 00:00:00
abstract：:The YUH1 gene coding for ubiquitin C-terminal hydrolase 1, a deubiquitinating enzyme, was cloned from the Saccharomyces cerevisiae genomic DNA and expressed in Escherichia coli. YUH1 was fused with the 6 histidine tag at the N-terminus (H6YUH1) or C-terminus (YUH1H6) and purified by an immobilized metal affinity chrom...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.07.005
更新日期：2007-11-01 00:00:00
abstract：:Autophagy is the process of degradation of intracellular proteins through the lysosome. Members of the tripartite motif (TRIM) proteins have shown to directly recognize autophagic cargo and also to act as a hub for the phagophore nucleation complex. The TRIM proteins are classically characterized by the presence of an...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2017.10.014
更新日期：2018-03-01 00:00:00
abstract：:Proteins are essential throughout the biological and biomedical sciences and the purification strategies of proteins of interest have advanced over centuries. Elastin-like polypeptides (ELPs) are compound polymers that have recently been highlighted for their sharp and reversible phase transition property when heated ...
journal_title：Protein expression and purification
pub_type： 杂志文章,评审
doi：10.1016/j.pep.2018.09.006
更新日期：2019-01-01 00:00:00
abstract：:Chinese hamster dihydrofolate reductase (ch-DHFR) was overexpressed in Escherichia coli DH5 alpha under the transcriptional control of PRPL promoters regulated by temperature-sensitive repressors. The desired recombinant product is soluble and constitutes about 30% of the total soluble proteins of the bacterial cell. ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1996.0676
更新日期：1997-03-01 00:00:00
abstract：:We report the molecular cloning, expression, and single-step homogeneous purification of a full-length asparaginyl tRNA synthetase (NRS) from Fasciola gigantica (FgNRS). Fasciola gigantica is a parasitic liver fluke of the class Trematoda. It causes fascioliasis that infects the liver of various mammals, including hum...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2017.10.009
更新日期：2018-03-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2008.12.012
更新日期：2009-05-01 00:00:00
abstract：:Antifreeze proteins (AFPs) enable organisms to survive under freezing or sub-freezing conditions. AFPs have a great potential in the low temperature storage of cells, tissues, organs, and foods. This process will require a large number of recombinant AFPs. In the present study, the recombinant carrot AFP was highly ex...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.01.019
更新日期：2004-06-01 00:00:00
abstract：:A highly efficient Escherichia coli expression system was established to obtain an appreciable quantity of antihypertensive peptide. The DNA-coding sequence for the Gly-Val-Tyr-Pro-His-Lys peptide was chemically synthesized and linked to form a ten-copy in tandem. It was cloned into the vector pET-15b and expressed in...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.05.001
更新日期：2015-09-01 00:00:00
