BacMam system for high-level expression of recombinant soluble and membrane glycoproteins for structural studies.

abstract：:Cell extracts of Clostridium kluyveri grown on ethanol plus succinate contained a NAD(H) dependent 4-hydroxybutanoate dehydrogenase (EC 1.1.1.61) at 66 U/mg. This enzyme was purified 42-fold under anaerobic conditions to homogeneity. Heat treatment, ion exchange chromatography on DEAE-cellulose, nondenaturing polyacry...

journal_title：Protein expression and purification

authors： Wolff RA,Kenealy WR

更新日期：1995-04-01 00:00:00

abstract：:An internal fragment (978 bp) corresponding to the dog zona pellucida glycoprotein-3 (DZP3), excluding the N-terminal signal sequence and the C-terminal transmembrane-like domain, was amplified by polymerase chain reaction from a full-length cDNA clone. The amplified SacI and PstI restricted fragment was cloned in-fra...

journal_title：Protein expression and purification

authors： Santhanam R,Panda AK,Kumar VS,Gupta SK

更新日期：1998-04-01 00:00:00

abstract：:We have constructed a plasmid, pAHOB1, with a 482-b AluI fragment containing the Escherichia coli glutaredoxin gene (grx) cloned under lambda PL promoter control. Growth of E. coli N4830/pAHOB1 cells at 30 degrees C followed by heat induction at 40 degrees C for 5 h resulted in expression of glutaredoxin as 20% of the...

journal_title：Protein expression and purification

authors： Björnberg O,Holmgren A

更新日期：1991-08-01 00:00:00

abstract：UNLABELLED:Recombinant protein production is a significant biotechnological process as it allows researchers to produce a specific protein in desired quantities. Escherichia coli (E. coli) is the most popular heterologous expression host for the production of recombinant proteins due to its advantages such as low cost,...

journal_title：Protein expression and purification

authors： Habibi N,Samian MR,Hashim SZ,Norouzi A

更新日期：2014-03-01 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：:Process development and optimization studies were performed in order to improve the purification process of (rhIFN-gamma). The objective was to generate material with higher purity and quantity. An in-process control screening was developed to obtain the optimal condition for column chromatographic purification by mea...

journal_title：Protein expression and purification

authors： Mohammadian-Mosaabadi J,Naderi-Manesh H,Maghsoudi N,Nassiri-Khalili MA,Masoumian MR,Malek-Sabet N

更新日期：2007-02-01 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

authors： Robinson RA,Burgess WH,Emerson SU,Leibowitz RS,Sosnovtseva SA,Tsarev S,Purcell RH

更新日期：1998-02-01 00:00:00

abstract：:alpha 1-Acid glycoprotein could be isolated by a one-step extraction method from human sera and plasma. Protein recovered in the water phase after extraction with phenol at 70 degrees C for 20 min was verified as human alpha 1-acid glycoprotein when it was compared with the reference standard human alpha 1-acid glycop...

journal_title：Protein expression and purification

authors： Chan J,Yu D

更新日期：1991-02-01 00:00:00

abstract：:The gene for the extremely thermophilic and thermostable 5'-methylthioadenosine phosphorylase from the archaeon Sulfolobus solfataricus was expressed at a high level in Escherichia coli thus providing a basis for detailed structural and functional studies of the enzyme. The recombinant enzyme was purified to homogenei...

journal_title：Protein expression and purification

authors： Cacciapuoti G,Fusco S,Caiazzo N,Zappia V,Porcelli M

更新日期：1999-06-01 00:00:00

abstract：:The entire extracellular domain of the human heat-stable enterotoxin (ST) receptor as well as a truncated N-terminal domain were cloned as glutathione S-transferase fusion proteins and expressed in Escherichia coli. The recombinant fusion proteins were purified from both the cytosol and the inclusion body fractions by...

journal_title：Protein expression and purification

authors： Nandi A,Mathew R,Visweswariah SS

更新日期：1996-09-01 00:00:00

abstract：:Characterizing protein complexes and identifying their subunits promote our understanding of the machinery involved in many in vivo processes. Proteomic studies can identify a protein's binding partners, and this can provide insight into how protein complexes function and how they are regulated. In addition, the compo...

journal_title：Protein expression and purification

authors： Banks CA,Kong SE,Washburn MP

更新日期：2012-12-01 00:00:00

abstract：:α-N-Acetylgalactosaminidase (α-GalNAc-ase; EC.3.2.1.49) is an exoglycosidase specific for the hydrolysis of terminal α-linked N-acetylgalactosamine in various sugar chains. The cDNA corresponding to the α-GalNAc-ase gene was cloned from Aspergillus niger, sequenced, and expressed in the yeast Saccharomyces cerevisiae....

journal_title：Protein expression and purification

authors： Mrázek H,Benada O,Man P,Vaněk O,Křen V,Bezouška K,Weignerová L

更新日期：2012-01-01 00:00:00

abstract：:Recombinant protein expression from lac derived promoters by the autoinduction regime is based on diauxic growth of Escherichia coli on glucose and lactose. Glycerol is used as a supporting carbon source during the lactose-induced expression. While this glycerol-based formulation usually provides high cell densities, ...

journal_title：Protein expression and purification

authors： Ukkonen K,Mayer S,Vasala A,Neubauer P

更新日期：2013-10-01 00:00:00

abstract：:This is the first report of an insect esterase efficiently expressed in the methylotrophic yeast Pichia pastoris (so far insect esterases have been produced only in the baculovirus system). Having isolated a Tribolium castaneum carboxylesterase cDNA (TCE), we were initially unable to express it in Escherichia coli or ...

journal_title：Protein expression and purification

authors： Delroisse JM,Dannau M,Gilsoul JJ,El Mejdoub T,Destain J,Portetelle D,Thonart P,Haubruge E,Vandenbol M

更新日期：2005-08-01 00:00:00

abstract：:We present a new streptavidin-binding peptide for both the purification and the detection of recombinant proteins. The peptide possesses nanomolar-affinity for streptavidin and therefore was termed Nano-tag. The Nano-tag(15) is 15 amino acids long and binds to streptavidin with a dissociation constant of 4 nM and the ...

journal_title：Protein expression and purification

authors： Lamla T,Erdmann VA

更新日期：2004-01-01 00:00:00

abstract：:Hydrophobic interaction high-performance liquid chromatography (HIC-HPLC) was utilized for the separation of native human antithrombin (AT) and a partially denaturated form of AT, known as the latent form (L-AT). The AT used in this study is commercially available (Atenativ, Pharmacia & Upjohn, Sweden) and contains al...

journal_title：Protein expression and purification

authors： Karlsson G,Winge S

更新日期：2001-02-01 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：:The numerous proteins occluded within the molluscan shell play a key role in the control of the mineralization process. Although extensively studied, these proteins are still poorly known, mainly because they are difficult to fractionate. In the present paper, we present, for the first time, a simple combined strategy...

journal_title：Protein expression and purification

authors： Marin F,Pereira L,Westbroek P

更新日期：2001-10-01 00:00:00

abstract：:The Ca2+-ATPase accounts for the majority of Ca2+ removed from the cytoplasm during cardiac muscle relaxation. The Ca2+-ATPase is regulated by phospholamban, a 52 amino acid phosphoprotein, which inhibits Ca2+-ATPase activity by decreasing the apparent affinity of the ATPase for Ca2+. To study the physical mechanism o...

journal_title：Protein expression and purification

authors： Waggoner JR,Huffman J,Griffith BN,Jones LR,Mahaney JE

更新日期：2004-03-01 00:00:00

abstract：:Pyrroline-5-carboxylate reductase (P5CR) catalyzes the reduction of Delta1-pyrroline-5-carboxylate (P5C) to proline with concomitant oxidation of NAD(P)H to NAD(P)(+). The enzymatic cycle between P5C and proline is very important in many physiological and pathological processes. Human P5CR was over-expressed in Escher...

journal_title：Protein expression and purification

authors： Meng Z,Lou Z,Liu Z,Hui D,Bartlam M,Rao Z

更新日期：2006-09-01 00:00:00

abstract：:This study describes comparison between IPTG and lactose induction on expression of caprine growth hormone (cGH), enhancing cell densities of Escherichia coli cultures and refolding the recombinant cGH, produced as inclusion bodies, to biologically active state. 2-3 times higher cell densities were obtained in shake f...

journal_title：Protein expression and purification

authors： Khan MA,Sadaf S,Sajjad M,Waheed Akhtar M

更新日期：2009-11-01 00:00:00

abstract：:The erm proteins confer resistance to the MLS (macrolide-lincosamide-streptogramin B) antibiotics in various microorganisms, including pathogens, through dimethylation of a single adenine residue (A2085: Bacillus subtilis coordinate) of the 23S rRNA to reduce the affinity of antibiotics, thereby enabling the cells to ...

journal_title：Protein expression and purification

authors： Jin HJ,Yang YD

更新日期：2002-06-01 00:00:00

abstract：:Receptor tyrosine kinase like orphan receptor 2 (ROR2) is a co-receptor for some Wnt proteins including Wnt5a that activate the noncanonical Wnt/planar cell polarity (PCP) signaling pathway. Upregulation of ROR2 is associated with several cancer forms. The extracellular region of ROR2, which contains an immunoglobulin...

journal_title：Protein expression and purification

authors： Li Y,Han X,Xu W,Rao Z,Li X

更新日期：2019-06-01 00:00:00

abstract：:Strategies for the expression of precursors of eukaryotic secreted proteins as part of fused proteins in Escherichia coli have been explored. A fusion protein with beta-galactosidase at the N-terminal end and honeybee prepromelittin at the C-terminal end (beta-gal-pM) was expressed in low amounts as a cleaved polypept...

journal_title：Protein expression and purification

authors： He M,Adcock I,Chapman D,Lucy J,Austen B

更新日期：1991-10-01 00:00:00

abstract：:An expression library was generated from a partial NcoI and HindIII digest of genomic DNA from the thermophilic bacterium, Bacillus stearothermophilus P1. The DNA fragments were cloned into the expression vector pQE-60 and transformed into Escherichia coli M15[EP4]. Sequence analysis of a lipase gene showed an open re...

journal_title：Protein expression and purification

authors： Sinchaikul S,Sookkheo B,Phutrakul S,Pan FM,Chen ST

更新日期：2001-08-01 00:00:00

abstract：:Tyrosine hydroxylase is the rate-limiting step in the synthesis of dopamine and is tightly regulated. Previous studies have shown it to be covalently modified and potently inhibited by 3,4-dihydroxyphenylacetaldehyde (DOPAL), an endogenous neurotoxin via dopamine catabolism which is relevant to Parkinson's disease. In...

journal_title：Protein expression and purification

authors： Higgins CA,Vermeer LM,Doorn JA,Roman DL

更新日期：2012-08-01 00:00:00

abstract：:The gene coding for phosphoglucomutase (PGM) from Oryctolagus cuniculus (rabbit) has been expressed in Escherichia coli under a T7 expression system with a His-tag. About half of the expressed PGM protein was present in inclusion bodies, but this protein was inactive when solubilized. The protein in the soluble cell f...

journal_title：Protein expression and purification

authors： Chae YK,Markley JL

更新日期：2000-10-01 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

authors： Ko JH,Hahm MS,Kang HA,Nam SW,Chung BH

更新日期：2002-08-01 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

authors： Xu L,Zhang C,Liu L,Zhang Y,Wang Q,Wang J,Liu Y,Su Z

更新日期：2017-11-01 00:00:00

abstract：:The heat shock protein hsp60 plays a functional role in insulin-dependent diabetes mellitus. The hsp60 epitope p277 (aa 437-aa 460) is effective in vaccinating mice against diabetes. A synthetic peptide gene (p277) that encodes the human hsp60 epitope was cloned to the 3' end of the cellulose-binding domain gene (cbd)...

journal_title：Protein expression and purification

authors： Shpigel E,Elias D,Cohen IR,Shoseyov O

更新日期：1998-11-01 00:00:00