Tabtoxin-resistant protein: overexpression, purification, and characterization.

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：:The availability of recombinant monomeric alkaline phosphatase (AP) is highly desirable in analytical applications involving AP fusion proteins. The cobalt-dependant alkaline phosphatase IV from Bacillus subtilis (BSAP), which was reported to be strongly monomeric, was overexpressed in Escherichia coli using pET autoi...

journal_title：Protein expression and purification

authors： Koksharov M,Lv C,Zhai X,Ugarova N,Huang E

更新日期：2013-08-01 00:00:00

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

authors： Hong SY,Kim TG,Kwon TH,Jang YS,Yang MS

更新日期：2007-07-01 00:00:00

abstract：:Soybean root nodules possess a developmentally regulated acid phosphatase (ACP) that exhibits the highest specificity for purine 5'-nucleoside monophosphates. The enzyme is a glycosylated dimer of 28- and 31-kDa subunits, which appear to be products of the same gene but differ in posttranslational modifications. In or...

journal_title：Protein expression and purification

authors： Penheiter AR,Klucas RV,Sarath G

更新日期：1998-10-01 00:00:00

abstract：:In plant, the first and the third steps of the synthesis of methionine and threonine are catalyzed by a bifunctional enzyme, aspartate kinase-homoserine dehydrogenase (AK-HSDH). In this study, we report the first purification and characterization of a highly active threonine-sensitive AK-HSDH from plants (Arabidopsis ...

journal_title：Protein expression and purification

authors： Paris S,Wessel PM,Dumas R

更新日期：2002-02-01 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

authors： Han ZS,Li QW,Zhang ZY,Xiao B,Gao DW,Wu SY,Li J,Zhao HW,Jiang ZL,Hu JH

更新日期：2007-05-01 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

authors： Favacho AR,Kurtenbach E,Sardi SI,Gouvea VS

更新日期：2006-04-01 00:00:00

abstract：:Recombinant barley high pI alpha-glucosidase was produced by high cell-density fermentation of Pichia pastoris expressing the cloned full-length gene. The gene was amplified from a genomic clone and exons (coding regions) were assembled by overlap PCR. The resulting cDNA was expressed under control of the alcohol oxid...

journal_title：Protein expression and purification

authors： Naested H,Kramhøft B,Lok F,Bojsen K,Yu S,Svensson B

更新日期：2006-03-01 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：:The HIV-1 Gag precursor protein, Pr55(Gag), is a multi-domain polyprotein that drives HIV-1 assembly. The morphological features of HIV-1 suggested Pr55(Gag) assumes a variety of different conformations during virion assembly and maturation, yet structural determination of HIV-1 Pr55(Gag) has not been possible due to ...

journal_title：Protein expression and purification

authors： McKinstry WJ,Hijnen M,Tanwar HS,Sparrow LG,Nagarajan S,Pham ST,Mak J

更新日期：2014-08-01 00:00:00

abstract：:Production of VCSM13 phage displaying a high density of CD147 ectodomain (CD147Ex) was achieved when culturing conditions were modulated. A phagemid expressing CD147Ex was constructed and used to produce phage display CD147Ex gpVIII fusion protein in TG1 Escherichia coli. Displaying of CD147Ex via gpVIII was successfu...

journal_title：Protein expression and purification

authors： Intasai N,Arooncharus P,Kasinrerk W,Tayapiwatana C

更新日期：2003-12-01 00:00:00

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 recomb...

journal_title：Protein expression and purification

authors： McCall EJ,Danielsson A,Hardern IM,Dartsch C,Hicks R,Wahlberg JM,Abbott WM

更新日期：2005-07-01 00:00:00

abstract：:A simple scheme for the rapid and efficient isolation of rat pro-atrial natriuretic factor (pro-ANF) has been developed. An isolated rat adrenal cell bioassay for ANF was established to optimize heart tissue extraction and chromatography conditions. This assay is based on the ability of ANF to inhibit angiotensin II-s...

journal_title：Protein expression and purification

authors： Belcourt D,Varma DR,Toney K,Bennett HP

更新日期：1990-09-01 00:00:00

abstract：:We have investigated the suitability of Pichia pastoris as an expression system for the candidate therapeutic protein, Sonic hedgehog fused to an immunoglobulin Fc domain (Shh-Fc). Sonic hedgehog is a morphogen protein involved in the patterning of a wide range of tissues during animal embryogenesis. The presence of S...

journal_title：Protein expression and purification

authors： Shapiro RI,Wen D,Levesque M,Hronowski X,Gill A,Garber EA,Galdes A,Strauch KL,Taylor FR

更新日期：2003-06-01 00:00:00

abstract：:Hydrophobic charge-induction chromatography (HCIC) using 4-mercaptoethylpyridine (4-MEP) as the ligand is used to purify antibodies. The 4-MEP resin ligand has high affinity for antibodies, which makes it difficult to optimize the elution conditions. Recent studies showed that arginine is effective at eluting and puri...

journal_title：Protein expression and purification

authors： Hirano A,Maruyama T,Shiraki K,Arakawa T,Kameda T

更新日期：2017-01-01 00:00:00

abstract：:HtrA2 is an apoptosis-activating protein to enhance the apoptotic process by preventing the formation of the IAP-caspase complex, thus freeing caspase to trigger the apoptosis pathway. Here, we presented the full-length sequence of HtrA2 from the black tiger shrimp (PmHtrA2). The full-length PmHtrA2 transcript was 140...

journal_title：Protein expression and purification

authors： Suwannaboon R,Phiwsaiya K,Senapin S,Khunrae P,Rattanarojpong T

更新日期：2013-12-01 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

authors： Krebs MP,Spudich EN,Spudich JL

更新日期：1995-12-01 00:00:00

abstract：:Deformylation of the initiator N-formylmethionine does not always proceed to completion for proteins overexpressed in Escherichia coli. To overcome this limitation, the def gene encoding the Escherichia coli peptide deformylase was cloned into the plysS plasmid under the tetracycline (Tc) promoter control. The efficie...

journal_title：Protein expression and purification

authors： Tang J,Hernández G,LeMaster DM

更新日期：2004-07-01 00:00:00

abstract：:A family of restriction enzyme- and ligation-independent cloning vectors has been developed for producing recombinant His-tagged fusion proteins in Escherichia coli. These are based on pURI2 and pURI3 expression vectors which have been previously used for the successful production of recombinant proteins at the millig...

journal_title：Protein expression and purification

authors： Curiel JA,de Las Rivas B,Mancheño JM,Muñoz R

更新日期：2011-03-01 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：:Recombinant protein expression in insect cells varies greatly from protein to protein. A fusion tag that is not only a tool for detection and purification, but also enhances expression and/or solubility would greatly facilitate both structure/function studies and therapeutic protein production. We have shown that fusi...

journal_title：Protein expression and purification

authors： Liu L,Spurrier J,Butt TR,Strickler JE

更新日期：2008-11-01 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

authors： li J,Li B,Li G,Ren J,Zhang J,Xu C,Yang X,Liu S,Fu L,Chen W

更新日期：2008-02-01 00:00:00

abstract：:The DEX gene encoding an extracellular dextranase from Lipomyces starkeyi was cloned into vector pPIC9k-His6 and was expressed in Pichia pastoris GS115 strain under the control of AOX1 promoter. After 107 h of the 5L-scaled fermentation, wet cells weight of the recombinant P. pastoris Mut(+) strain reached to 588.6g/L...

journal_title：Protein expression and purification

authors： Chen L,Zhou X,Fan W,Zhang Y

更新日期：2008-03-01 00:00:00

abstract：:Baculovirus mediated gene transduction of mammalian cells (BacMam) is an emerging technique for rapid recombinant protein expression in mammalian cells. We constructed two baculovirus transfer vectors that incorporate several mammalian transcriptional regulatory elements necessary for high-level protein expression in ...

journal_title：Protein expression and purification

authors： Dukkipati A,Park HH,Waghray D,Fischer S,Garcia KC

更新日期：2008-12-01 00:00:00

abstract：:Soluble guanylate cyclase (sGC) has been purified from 100 L cell culture infected by baculovirus using the newer and highly effective titerless infected-cells preservation and scale-up (TIPS) method. Successive passage of the enzyme through DEAE, Ni(2+)-NTA, and POROS Q columns obtained approximately 100mg of protein...

journal_title：Protein expression and purification

authors： Emmons TL,Mathis KJ,Shuck ME,Reitz BA,Curran DF,Walker MC,Leone JW,Day JE,Bienkowski MJ,Fischer HD,Tomasselli AG

更新日期：2009-06-01 00:00:00

abstract：:3-Deoxy-d-manno-octulosonate 8-phosphate synthase (KDO8PS) [EC 4.1.2.16] is the first and rate-limiting enzyme in the 3-deoxy-d-manno-octulosonate (KDO) biosynthetic pathway. The enzyme is widely expressed in bacteria and plants. Their well conserved protein sequences imply a similar oligomeric arrangement. However, t...

journal_title：Protein expression and purification

authors： Zhang F,Xu Y,Zhang Z

更新日期：2014-09-01 00:00:00

abstract：:A baculovirus vector system that expresses cloned DNA sequences as glutathione S-transferase fusion proteins was developed. This system was used to express and purify the lymphocyte-specific tyrosine kinase p56lck. This recombinant p56lck was purified to homogeneity in a single chromatography step using glutathione re...

journal_title：Protein expression and purification

authors： Spana C,O'Rourke EC,Bolen JB,Fargnoli J

更新日期：1993-10-01 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

authors： Tomisawa S,Sato Y,Kamiya M,Kumaki Y,Kikukawa T,Kawano K,Demura M,Nakamura K,Ayabe T,Aizawa T

更新日期：2015-08-01 00:00:00

abstract：:To obtain strong bacterial expression of proteins that seem to be hard to express in bacteria or are highly toxic for bacteria, it is possible to create a palette of similar constructs, differing only by several nucleotides, gradually deleted from the full-length clone by exonuclease III. When a construct is equipped ...

journal_title：Protein expression and purification

authors： Pecenková T,Filigarová M,Cerovská N

更新日期：2005-05-01 00:00:00

abstract：:A process for bacterial expression and purification of the recombinant major wasp allergen Antigen 5 (Ves v 5) was developed to produce protein for diagnostic and therapeutic applications for type 1 allergic diseases. Special attention was focused on medium selection, fermentation conditions, and efficient refolding p...

journal_title：Protein expression and purification

authors： Kischnick S,Weber B,Verdino P,Keller W,Sanders EA,Anspach FB,Fiebig H,Cromwell O,Suck R

更新日期：2006-06-01 00:00:00