Abstract:
:Anthrax toxin consists of three separate proteins, protective antigen (PA), lethal factor (LF), and edema factor (EF). PA binds to the receptor on mammalian cells and facilitates translocation of EF or LF into the cytosol. PA is the primary component of several anthrax vaccines. In this study we expressed and purified PA from Escherichia coli. The purification of PA from E. coli was possible after transporting the protein into the periplasmic space using the outer membrane protein A signal sequence. The purification involved sequential chromatography through hydroxyapatite, DEAE Sepharose CL-4B, followed by Sephadex G-100. The typical yield of purified PA from this procedure was 500 microg/liter. PA expressed and purified from E. coli was similar to the PA purified from Bacillus anthracis in its ability to lyse a macrophage cell line (J774A.1). The present results suggest that a signal sequence is required for the efficient translocation of PA into E. coli periplasmic space.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Sharma M,Swain PK,Chopra AP,Chaudhary VK,Singh Ydoi
10.1006/prep.1996.0005subject
Has Abstractpub_date
1996-02-01 00:00:00pages
33-8issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(96)90005-4journal_volume
7pub_type
杂志文章abstract::Hepatitis C virus encodes two enveloped glycoproteins, E1 and E2, which are involved in viral attachment and entry into target cells. We have obtained in insect cells infected by recombinant baculovirus a chimeric secreted recombinant protein, E1(341)E2(661,) containing the ectodomains of E1 and E2. The described proc...
journal_title:Protein expression and purification
pub_type: 杂志文章
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更新日期:2010-06-01 00:00:00
abstract::Protein convertase 1/3 is a serine endoproteinase present in the regulated secretory pathway of endocrine and neuroendocrine cells. It is responsible for the processing of numerous prohormones and proneuropeptides into their biologically active moieties, often following cleavage at pairs of basic residues. The determi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.06.014
更新日期:2004-10-01 00:00:00
abstract::Homocysteine alpha,gamma-lyase from the anaerobic protozoan parasite Trichomonas vaginalis has been cloned from genomic DNA using PCR methods and expressed in Escherichia coli with a vector containing the T7 promoter. The recombinant homocysteine alpha,gamma-lyase (rHCYase) is expressed as the major protein in the hos...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1998.0955
更新日期:1998-11-01 00:00:00
abstract::Whey acidic proteins (WAP) belong to a large gene family of antibacterial peptides that perform critical immune system functions. The function of human epididymis protein 4 (HE4), a 124-amino acid long polypeptide that has two whey acidic protein four-disulfide core (WFDC) domains, is not well studied. Here, a fusion ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.08.004
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abstract::NAD(+)-dependent malic enzyme (NAD-ME) gene from Escherichia coli K12 was inserted into an expression vector pET24b(+) and transformed into E. coli BL21 (DE3). Recombinant NAD-ME was expressed upon IPTG induction, purified with affinity chromatography, and biochemically characterized. The results showed that recombina...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.11.017
更新日期:2007-05-01 00:00:00
abstract::Pullulanases are well-known starch-debranching enzymes that are widely used for hydrolysis of a-1,6-glycosidic linkages in starch, pullulan, amylopectin, and other oligosaccharides. Escherichia coli is a popular heterologous expression host for generating target enzymes. However, cells have to be disrupted to obtain t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2018.09.011
更新日期:2019-03-01 00:00:00
abstract::Plasmids pQE-60 and pQE-30 containing 6 x His-tag sequence were used for expression of fragments of HCV structural and non-structural proteins in Escherichia coli (E. coli). The following fragments were used: core (1-98 aa), NS3 (202-482 aa), and tetramer of hypervariable region 1 (HVR1) of E2 protein. The constructed...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.06.011
更新日期:2006-11-01 00:00:00
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 chromatogra...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.10.011
更新日期:2008-02-01 00:00:00
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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::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
pub_type: 杂志文章
doi:10.1016/j.pep.2008.02.012
更新日期:2008-06-01 00:00:00
abstract::Vascular leak syndrome (VLS) is the major dose-limiting toxicity of immunotoxin therapy. In our previous study, a modified PE38KDEL, denoted PE38KDELKQK, was engineered to eliminate VLS. The PE38KDELKQK-based immunotoxin has been proved to retain potent anti-tumor activity but with a remarkable attenuation in VLS. In ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.09.009
更新日期:2008-03-01 00:00:00
abstract::The gene encoding glucose oxidase (GOD) from Aspergillus niger was expressed as a secretory product in the yeast Saccharomyces cerevisiae. Six consecutive histidine residues were fused to the C-terminus of GOD to facilitate purification. The recombinant GOD-His(6) secreted by S. cerevisiae migrated as a broad diffuse ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00035-9
更新日期:2002-08-01 00:00:00
abstract::A full-length cDNA sequence of plant type CRY (designated Hae-P-CRY) was cloned from the green alga Haematococcus pluvialis. The cDNA sequence was 3608 base pairs (bp) in length, which contained a 2988-bp open reading frame encoding 995 amino acids with molecular mass of 107.7 kDa and isoelectric point of 6.19. Multip...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105633
更新日期:2020-08-01 00:00:00
abstract::Ciliary neurotrophic factor (CNTF) is a promising candidate for the treatment of neurodegenerative or metabolic diseases, but suffers rapid clearance in body. Herein we constructed a new long-acting recombinant human CNTF (rhCNTF) by genetic fusion with an albumin-binding domain (ABD) through a flexible peptide linker...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2017.07.006
更新日期:2017-11-01 00:00:00
abstract::Escherichia coli cells were transformed with an expression vector constructed by inserting a DNA fragment encoding a Kazal-type trypsin inhibitor from mouse seminal vesicle into pGEX-2. The cloned cells were able to produce a high yield of a chimeric polypeptide made by fusing the trypsin inhibitor to glutathione S-tr...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1994.1003
更新日期:1994-02-01 00:00:00
abstract::Nedd4 is an E3 ubiquitin ligase that has received increased attention due to its role in the maintenance of proteostasis and in cellular stress responses. Investigation of Nedd4 enzymology has revealed a complex enzymatic mechanism that involves intermolecular interactions with upstream E2 conjugating enzymes and with...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105649
更新日期:2020-09-01 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::The Human Secretome Project aims to produce and purify all human secreted proteins as full-length. In order to enable this, a robust, gentle and effective purification process is needed, where multiple proteins can be purified in parallel. For this reason, a purification system based on a Protein C-tag and the HPC4 an...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105698
更新日期:2020-11-01 00:00:00
abstract::Integrated studies that address proteins structure and function in the new era of systems biology and genomics often require the application of high-throughput approaches for parallel production of many different purified proteins from the same organism. Cytochromes c-electron transfer proteins carrying one or more he...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.06.014
更新日期:2008-11-01 00:00:00
abstract::We have used Ni(2+)-affinity chromatography as a rapid and efficient method to purify a sensory rhodopsin I (SR-I) derivative containing six consecutive histidine residues at its C-terminus (His-tagged SR-I). The protein was expressed in Halobacterium salinarium by integrating the corresponding gene at the chromosomal...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1995.0009
更新日期:1995-12-01 00:00:00
abstract::Protein aggregation is a main barrier hindering structural and functional studies of a number of interesting biological targets. The E6 oncoprotein of Human Papillomavirus strain 16 (E6(16)) is difficult to express under a native soluble form in bacteria. Produced as an unfused sequence, it forms inclusion bodies. Fus...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.07.029
更新日期:2007-01-01 00:00:00
abstract::We have previously reported that a variety of solid human tumor cell lines express a large number of receptors for interleukin-13 (IL-13). These receptors could be targeted with a chimeric fusion protein consisting of human IL-13 and a truncated form of Pseudomonas exotoxin (PE). We describe here optimization of criti...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.10.012
更新日期:2005-02-01 00:00:00
abstract::Mannan outer chain N-glycan structures are yeast/fungal-specific typically found on secreted and cell wall glycoproteins. Mannan outer chains consist of an alpha-1,6 polymannose backbone attached to a Man(8-10)(GlcNAc)(2) core. The backbone contains branches of alpha-1,2 mannose residues, terminated with alpha-1,3 man...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.02.013
更新日期:2009-07-01 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::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::Arginase (EC 3.5.3.1; L-arginine amidinohydrolase) is a key enzyme of the urea cycle that catalyses the conversion of arginine to ornithine and urea, which is the final cytosolic reaction of urea formation in the mammalian liver. The recombinant strain of the yeast Saccharomyces cerevisiae that is capable of overprodu...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.09.001
更新日期:2012-01-01 00:00:00
abstract::The metalloprotease PrtV from Vibrio cholerae serves an important function for the bacteria's ability to invade the mammalian host cell. The protein belongs to the family of M6 proteases, with a characteristic zinc ion in the catalytic active site. PrtV constitutes a 918 amino acids (102 kDa) multidomain pre-pro-prote...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.01.012
更新日期:2014-04-01 00:00:00
abstract::The expression of human lactoferrin in the mammary gland is an attractive approach to diminish its current production cost. Previous attempts to produce lactorferrin in the milk of transgenic animals resulted in very high cost and uncertain results. In this paper, we have directly infused replication-defective adenovi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.11.019
更新日期:2007-05-01 00:00:00
abstract::A highly efficient cell-free translation system has been combined with suppressor tRNA technology to substitute nor-Tyr and 3-fluoro-Tyr in place of Tyr183 at the DNA polymerase active site of p66 of human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT). Supplementing the wild-type HIV-1 p51 RT subunit ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.07.019
更新日期:2004-11-01 00:00:00
abstract::The cofactor heme (Fe-protoporphyrin IX) plays many important roles in biology. Identification of novel proteins for the transport, chaperoning and delivery of heme in cells is of widespread interest. Here, we describe the use of heme conjugated magnetic beads for the isolation of heme-binding proteins from complex pr...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.10.001
更新日期:2011-03-01 00:00:00