High-level over-expression, purification, and crystallization of a novel phospholipase C/sphingomyelinase from Pseudomonas aeruginosa.

abstract：:Stem cell factor (SCF) known as the c-kit ligand is a two disulfide bridge-containing cytokine in the regulation of the development and function of hematopoietic cell lineages and other cells such as mast cells, germ cells, and melanocytes. The secreted soluble form of SCF exists as noncovalently associated homodimer ...

journal_title：Protein expression and purification

authors： Akuta T,Kikuchi-Ueda T,Imaizumi K,Oshikane H,Nakaki T,Okada Y,Sultana S,Kobayashi K,Kiyokawa N,Ono Y

更新日期：2015-01-01 00:00:00

abstract：:Proteomics has become a major focus as researchers attempt to understand the vast amount of genomic information. Protein complexity makes identifying and understanding gene function inherently difficult. The challenge of studying proteins in a global way is driving the development of new technologies for systematic an...

journal_title：Protein expression and purification

authors： Lesley SA

更新日期：2001-07-01 00:00:00

abstract：:Fab fragments isolated from papain digests of monoclonal antibodies have a wide variety of uses in analytical and in both in vivo and in vitro diagnostic applications. A novel, non-affinity method which uses the Gradiflow to purify Fab fragments from the papain digest of a mouse IgG1 anti-c-myc monoclonal antibody is ...

journal_title：Protein expression and purification

authors： Coleman L,Mahler SM

更新日期：2003-12-01 00:00:00

abstract：:DNA gyrase is a type IIA topoisomerase found in bacteria but not in humans. The enzyme is required for bacterial DNA replication and transcription, and is an important antibacterial target that is sensitive to the widely-used fluoroquinolone drugs. Due to the emergence of fluoroquinolone resistance, the discovery of n...

journal_title：Protein expression and purification

authors： Aung HL,Samaranayaka CU,Enright R,Beggs KT,Monk BC

更新日期：2015-03-01 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

authors： Kalbina I,Lagerqvist N,Moiane B,Ahlm C,Andersson S,Strid Å,Falk KI

更新日期：2016-11-01 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

authors： Yu HA,Kim SG,Kim EJ,Lee WJ,Kim DO,Park K,Park YC,Seo JH

更新日期：2007-11-01 00:00:00

abstract：:The transcription factor Sp1 is a regulator of TATA-less genes. It belongs to the Cys₂-His₂ zinc finger domain-containing family. A zebrafish cDNA encoding a peptide homologous to mammalian Sp1 was cloned and inserted into a pET43.1a vector and expressed in Escherichia coli Rosetta (DE3) cells as a Nus-His-tag fusion ...

journal_title：Protein expression and purification

authors： Lin CJ,Hsiao TH,Chung YS,Chang WN,Yeh TM,Chen BH,Fu TF

更新日期：2011-03-01 00:00:00

abstract：:Azotobacter vinelandii has recently been used for a variety of genetic experiments which take advantage of its facile transformation system and its high-frequency homologous recombination. One gene that has been cloned and sequenced is the fdxA gene that encodes a small Fe-S protein called A. vinelandii ferredoxin I (...

journal_title：Protein expression and purification

authors： Vázquez A,Shen B,Negaard K,Iismaa S,Burgess B

更新日期：1994-02-01 00:00:00

abstract：:Human pancreatic ribonuclease (HP-RNase) has considerable promise as a therapeutic agent. Structure-function analyses of HP-RNase have been impeded by the difficulty of obtaining the enzyme from its host. Here, a gene encoding HP-RNase was designed, synthesized, and inserted into two expression vectors that then direc...

journal_title：Protein expression and purification

authors： Ribó M,delCardayré SB,Raines RT,de Llorens R,Cuchillo CM

更新日期：1996-05-01 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

authors： Thomson CA,Ananthanarayanan VS

更新日期：2001-10-01 00:00:00

abstract：:Heparinase I (HepA) was originally isolated from Flavobacterium heparinum (F. heparinum) and specifically cleaves heparin/heparan sulfate in a site-dependent manner, showing great promise for producing low molecular weight heparin (LMWH). However, expressing recombinant HepA is extremely difficult in Escherichia coli ...

journal_title：Protein expression and purification

authors： Huang J,Cao L,Guo W,Yuan R,Jia Z,Huang K

更新日期：2012-06-01 00:00:00

abstract：:Lactase deficiency problem discourages many adults from consuming milk as a major source of micro- and macronutrients. Enzymatic hydrolysis of lactose is an ideal solution for this problem but such processing adds significant costs. In this study, a cold active β-galactosidase from Planococcus sp-L4 (bgal) was optimiz...

journal_title：Protein expression and purification

authors： Mahdian SM,Karimi E,Tanipour MH,Parizadeh SM,Ghayour-Mobarhan M,Bazaz MM,Mashkani B

更新日期：2016-09-01 00:00:00

abstract：:The covalent addition of 4-amino-4-deoxy-L-arabinose (L-Ara4N) groups to lipid A, which resides in the outer membranes of bacteria such as Salmonella typhimurium and Escherichia coli, is the final step in the polymyxin-resistance pathway in these organisms. This modification is catalyzed by the inner membrane protein ...

journal_title：Protein expression and purification

authors： Bretscher LE,Morrell MT,Funk AL,Klug CS

更新日期：2006-03-01 00:00:00

abstract：:Recombinant cytochrome P450 (CYP or P450) enzymes are useful for drug metabolism research and thereby many expression and purification systems have been developed. Here, we provide a method for the purification of human P450s 3A4 and 1A2 expressed in Escherichia coli using mixed micelles containing anionic phospholipi...

journal_title：Protein expression and purification

authors： Ahn T,Bae CS,Yun CH

更新日期：2014-09-01 00:00:00

abstract：:Reticulons are a large family of integral membrane proteins that are ubiquitous in eukaryotes and play a key role in functional remodelling of the endoplasmic reticulum membrane. The reticulon family is especially large in plants, with the Arabidopsis thaliana genome containing twenty-one isoforms. Reticulons vary in ...

journal_title：Protein expression and purification

authors： Chow M,Sklepari M,Frigerio L,Dixon AM

更新日期：2018-12-01 00:00:00

abstract：:p53 protein is an important regulation factor that can bind to p53 mRNA to regulate its translation in human and murine. To determine if a similar interaction exists in zebrafish and if the interaction affects zebrafish development, we cloned and expressed p53 protein from zebrafish in Escherichia coli. Soluble p53 pr...

journal_title：Protein expression and purification

authors： Zhao X,Liu M,Wu N,Ding L,Liu H,Lin X

更新日期：2010-08-01 00:00:00

abstract：:High-level extracellular production of Fusarium solani cutinase was achieved using a Pichia pastoris expression system. The cutinase-encoding gene was cloned into pPICZalphaA with the Saccharomyces cerevisiae alpha-factor signal sequence and methanol-inducible alcohol oxidase promoter by two different ways. The additi...

journal_title：Protein expression and purification

authors： Kwon MA,Kim HS,Yang TH,Song BK,Song JK

更新日期：2009-11-01 00:00:00

abstract：:A new procedure for the large-scale purification of the recombinant thermostable chitinase (Chi40) cloned from Streptomyces thermoviolaceus in various expression vectors in Escherichia coli is described. Chi40 was overproduced in the cytosolic and secreted forms. The cytosolic form (Chi40c) was highly overproduced and...

journal_title：Protein expression and purification

authors： Christodoulou E,Duffner F,Vorgias CE

更新日期：2001-10-01 00:00:00

abstract：:In this work, we present the production of an active 43 aa recombinant human beta-defensin-1 (rhBD-1(43)) in Escherichia coli AD202 cells using specific pLMM1-rhBD-1 expression system. Unique solubility properties of the C-terminal fragment of light meromyosin (LMM) allowed us to overcome foreseeable problems with iso...

journal_title：Protein expression and purification

authors： Cipáková I,Hostinová E,Gasperík J,Velebný V

更新日期：2004-09-01 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

authors： Woo JH,Liu YY,Mathias A,Stavrou S,Wang Z,Thompson J,Neville DM Jr

更新日期：2002-07-01 00:00:00

abstract：:Antibodies specific to β-Glucocerebrosidase were selected from phage displayed naïve scFv libraries. Biopannings were performed against recombinant human protein β-Glucocerebrosidase immobilized on polystyrene surface, specific phages were eluted with 50% ethylene glycol in citrate buffer (pH 6.0). Several specific bi...

journal_title：Protein expression and purification

authors： Anisimov RL,Ershova OA,Ershov AV,Filatova MA,Katorkin SA,Simonov VM

更新日期：2020-06-01 00:00:00

abstract：:The superoxide dismutases (EC 1.15.1.1) are a family of enzymes that catalyze the dismutation of superoxide radical anion to dioxygen and hydrogen peroxide. The active site contains a critical metal ion such as manganese, iron, or copper. The copper-containing protein also has one zinc ion bound per subunit. The stand...

journal_title：Protein expression and purification

authors： Sutter B,Bounds PL,Koppenol WH

更新日期：2000-06-01 00:00:00

abstract：:Campylobacteriosis is a disease in humans caused by the infection from Campylobacter spp. Human cases are mainly due to Campylobacter jejuni, although C. coli can cause gastroenteritis in humans as well. The bacteria are commensal in chicken tract and can be contaminated into chicken products during processing. Obviou...

journal_title：Protein expression and purification

authors： Wenbap P,Seetang-Nun Y,Luangtongkum T,Khunrae P,Tuitemwong P,Rattanarojpong T

更新日期：2021-04-01 00:00:00

abstract：:We describe a single method to purify milligram amounts of authentic wild-type and mutant HIV-1 and HIV-2 Tat proteins overexpressed in Escherichia coli. This method takes advantage of the highly basic, positively charged RNA binding domain present in both HIV-1 and HIV-2 Tat, which also facilitated purification of HI...

journal_title：Protein expression and purification

authors： Orsini MJ,García-Martínez LF,Mavankal G,Gaynor RB,Debouck CM

更新日期：1996-09-01 00:00:00

abstract：:A fibrinolytic enzyme was purified from the cultured mycelia of Armillaria mellea by ion-exchange chromatography followed by gel filtration, and was designated A. mellea metalloprotease (AMMP). The purification protocol resulted in a 627-fold purification of the enzyme, with a final yield of 6.05%. The apparent molecu...

journal_title：Protein expression and purification

authors： Lee SY,Kim JS,Kim JE,Sapkota K,Shen MH,Kim S,Chun HS,Yoo JC,Choi HS,Kim MK,Kim SJ

更新日期：2005-09-01 00:00:00

abstract：:Heterologous protein expression in Escherichia coli is commonly used to obtain recombinant proteins for a variety of downstream applications. However, many proteins are not, or are only poorly, expressed in soluble form. High level expression often leads to the formation of inclusion bodies and an inactive product tha...

journal_title：Protein expression and purification

authors： Pacheco B,Crombet L,Loppnau P,Cossar D

更新日期：2012-01-01 00:00:00

abstract：:The P(II) proteins comprise a family of widely distributed signal transduction proteins that integrate the signals of cellular nitrogen, carbon and energy status, and then regulate, by protein-protein interaction, the activity of a variety of target proteins including enzymes, transcriptional regulators and membrane t...

journal_title：Protein expression and purification

authors： Moure VR,Razzera G,Araújo LM,Oliveira MAS,Gerhardt ECM,Müller-Santos M,Almeida F,Pedrosa FO,Valente AP,Souza EM,Huergo LF

更新日期：2012-01-01 00:00:00

abstract：:Botulinum neurotoxin serotype A (BoNT/A) is an extremely potent bacterial protein toxin. The Hc fragment of BoNT/A (AHc) was shown to be non-toxic, antigenic, and capable of eliciting a protective immunity in animals challenged with homologous BoNT. In this study, we synthesized AHc gene by using T4 DNA ligase and PCR...

journal_title：Protein expression and purification

authors： Chen R,Shi J,Cai K,Tu W,Hou X,Liu H,Xiao L,Wang Q,Tang Y,Wang H

更新日期：2010-05-01 00:00:00

abstract：:A crude cell extract from yeast Saccharomyces cerevisae was fractionated by affinity chromatography using the leucine tRNA gene as the recognition site. This approach enables the rapid purification of a protein, which retained its full DNA binding capacity during the enrichment procedure. The active fraction contains ...

journal_title：Protein expression and purification

authors： Kaufmann E

更新日期：1990-11-01 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

authors： Kumar L,Colomb W,Czerski J,Cox CR,Sarkar SK

更新日期：2018-08-01 00:00:00