Recovery of recombinant zebrafish p53 protein from inclusion bodies and its binding activity to p53 mRNA in vitro.

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 protein with high purity was successfully obtained using the optimized renaturation approach. Results of a UV-crosslinking experiment and immunoprecipitation:RT-PCR analysis confirmed that the purified p53 protein could bind specifically to its cognate mRNA. Our results suggest that selecting a suitable buffer is important for renaturing p53 protein from inclusion bodies. We also demonstrated a specific interaction between p53 and it own RNA in zebrafish. Measurement of the binding activity may be a useful approach for identifying the activity of recombinant p53 protein in vitro.

journal_name

Protein Expr Purif

authors

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

doi

10.1016/j.pep.2010.03.028

subject

Has Abstract

pub_date

2010-08-01 00:00:00

pages

262-6

issue

2

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(10)00103-8

journal_volume

72

pub_type

杂志文章
  • Heterologous protein production in Escherichia coli using the propionate-inducible pPro system by conventional and auto-induction methods.

    abstract::We examined expression of two plant genes encoding coclaurine N-methyltransferase (CMT) and norcoclaurine synthase (NCS) in Escherichia coli from the Salmonella entericaprpBCDE promoter (P(prpB)) and compared it to that from the strongest IPTG-inducible promoter, P(T7). In contrast to our previous study showing slight...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2008.06.008

    authors: Lee SK,Keasling JD

    更新日期:2008-10-01 00:00:00

  • Soybean disease resistance protein RHG1-LRR domain expressed, purified and refolded from Escherichia coli inclusion bodies: preparation for a functional analysis.

    abstract::Introduction and expression of foreign genes in bacteria often results accumulation of the foreign protein(s) in inclusion bodies (IBs). The subsequent processes of refolding are slow, difficult and often fail to yield significant amounts of folded protein. RHG1 encoded by rhg1 was a soybean (Glycine max L. Merr.) tra...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2006.12.017

    authors: Afzal AJ,Lightfoot DA

    更新日期:2007-06-01 00:00:00

  • Improved soluble expression of a single-chain antibody fragment in E. coli for targeting CA125 in epithelial ovarian cancer.

    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

    authors: Sharma SK,Suresh MR,Wuest FR

    更新日期:2014-10-01 00:00:00

  • High-level expression of human lactoferrin in the milk of goats by using replication-defective adenoviral vectors.

    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

    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

  • Characterization of ubiquitin C-terminal hydrolase 1 (YUH1) from Saccharomyces cerevisiae expressed in recombinant Escherichia coli.

    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

    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

  • Systematic optimization of expression and refolding of the Plasmodium falciparum cysteine protease falcipain-2.

    abstract::The Plasmodium falciparum cysteine protease falcipain-2 is a potential new target for antimalarial chemotherapy. In order to obtain large quantities of active falcipain-2 for biochemical and structural analysis, a systematic assessment of optimal parameters for the expression and refolding of the protease was carried ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.2001.1416

    authors: Sijwali PS,Brinen LS,Rosenthal PJ

    更新日期:2001-06-01 00:00:00

  • Purification of granulin-like polypeptide from the blood-sucking leech, Hirudo nipponia.

    abstract::A cysteine-rich (approximately 20%), low molecular weight (MW 6 kDa) polypeptide has been isolated from the Korean blood-sucking leech, Hirudo nipponia. From its amino acid composition and N-terminal amino acid sequence analysis, the new protein is similar to granulin (or epithelin), and so it has been named leech gra...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1999.1077

    authors: Hong SJ,Kang KW

    更新日期:1999-07-01 00:00:00

  • Overproduction and purification of sigmaS, the Escherichia coli stationary phase specific sigma transcription factor.

    abstract::This paper reports the overproduction and the details of a rapid method to purify active sigmaS monomers from a T7 RNA polymerase-based protein expression system. This 2-day procedure involves solubilizing inclusion bodies in sarkosyl detergent, removal of sarkosyl by dialysis, and a single gel filtration column chrom...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1996.0069

    authors: Nguyen LH,Burgess RR

    更新日期:1996-08-01 00:00:00

  • Efficient recovery of the functional IP10-scFv fusion protein from inclusion bodies with an on-column refolding system.

    abstract::A functional IP10-scFv fusion protein retaining the antibody specificity for acidic isoferritin and chemokine function was produced at high level in Esherichia coli (E. coli). IP10-scFv gene from the recombinant plasmid pc3IP104c9 was subcloned into pET28a fused to N-terminal His-tag sequence in frame and overexpresse...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2005.05.016

    authors: Guo JQ,Li QM,Zhou JY,Zhang GP,Yang YY,Xing GX,Zhao D,You SY,Zhang CY

    更新日期:2006-01-01 00:00:00

  • Purification and characterization of recombinant forms of murine Tcl1 proteins.

    abstract::The TCL1 gene, which is located on chromosome 14, plays a major role in human hematopoietic malignancies and encodes a 14-kDa protein whose function has not been determined. This gene is expressed in pre-B cells, in immature thymocytes, and, at low levels, in activated T cells but not in peripheral mature B cells and ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1999.1186

    authors: Du Bois GC,Song SP,Kulikovskaya I,Rothstein JL,Germann MW,Croce CM

    更新日期:2000-04-01 00:00:00

  • A simple and efficient method for generating high-quality recombinant Mical enzyme for in vitro assays.

    abstract::We have recently identified a new family of multidomain oxidoreductase (redox) enzymes, the MICALs, that directly regulate the actin cytoskeletal elements necessary for the morphology, motility, and trajectory of cells. Our genetic assays reveal that Mical is both necessary and sufficient for actin organization and ce...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2016.05.008

    authors: Wu H,Hung RJ,Terman JR

    更新日期:2016-11-01 00:00:00

  • Heme content of recombinant catalase from Psychrobacter sp. T-3 altered by host Escherichia coli cell growth conditions.

    abstract::The catalase gene of Psychrobacter sp. T-3 was cloned, and the gene product (PktA) was overexpressed in Escherichia coli. The specific activity of the purified PktA was slightly lower than that of the native purified enzyme obtained from Psychrobacter sp. T-3. Spectrophotometric measurements of the purified enzymes su...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2008.03.016

    authors: Kimoto H,Matsuyama H,Yumoto I,Yoshimune K

    更新日期:2008-06-01 00:00:00

  • Overexpression of the Bacillus subtilis and circulans xylanases in Escherichia coli.

    abstract::An efficient expression system for a low-molecular mass xylanase in Escherichia coli has been developed. A gene encoding the mature Bacillus circulans (Bc) xylanase was designed to imitate the frequency of degenerate codons used in E. coli. Appropriate degenerate codons were used to create multiple unique restriction ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1993.1026

    authors: Sung WL,Luk CK,Zahab DM,Wakarchuk W

    更新日期:1993-06-01 00:00:00

  • Cloning, expression, purification, and biochemical characterisation of the FIC motif containing protein of Mycobacterium tuberculosis.

    abstract::The role of FIC (Filamentation induced by cAMP)(2) domain containing proteins in the regulation of many vital pathways, mostly through the transfer of NMPs from NTPs to specific target proteins (NMPylation), in microorganisms, higher eukaryotes, and plants is emerging. The identity and function of FIC domain containin...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2012.08.020

    authors: Mishra S,Bhagavat R,Chandra N,Vijayarangan N,Rajeswari H,Ajitkumar P

    更新日期:2012-11-01 00:00:00

  • Expression and properties of arginyl-tRNA synthetase from jack bean (Canavalia ensiformis).

    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

    authors: Hogg J,Schiefermayr E,Schiltz E,Igloi GL

    更新日期:2008-10-01 00:00:00

  • High-level extracellular production of Rhizopus oryzae lipase in Pichia pastoris via a strategy combining optimization of gene-copy number with co-expression of ERAD-related proteins.

    abstract::Rhizopus oryzae lipase (ROL) is an important industrial enzyme limited in application due to its low production in native strains. Here, we used a new combined strategy to overexpress ROL in Pichia pastoris. An efficient method based on bio-brick was developed to construct a series of vectors harboring different copy ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2018.02.005

    authors: Jiao L,Zhou Q,Su Z,Xu L,Yan Y

    更新日期:2018-07-01 00:00:00

  • Expression of Sonic hedgehog-Fc fusion protein in Pichia pastoris. Identification and control of post-translational, chemical, and proteolytic modifications.

    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

    pub_type: 杂志文章

    doi:10.1016/s1046-5928(03)00062-7

    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

  • Purification, characterization, and crystallization of human pyrroline-5-carboxylate reductase.

    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

    pub_type: 杂志文章

    doi:10.1016/j.pep.2006.02.019

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

    更新日期:2006-09-01 00:00:00

  • Amino-terminal charge affects the periplasmic accumulation of recombinant heregulin/EGF hybrids exported using the Escherichia coli alkaline phosphatase signal sequence.

    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

    authors: Campion SR,Elsasser E,Chung R

    更新日期:1997-08-01 00:00:00

  • A new protocol for high-yield purification of recombinant human CXCL8((3-72))K11R/G31P expressed in Escherichia coli.

    abstract::The ELR-CXC chemokines are important to neutrophil inflammation in many acute and chronic diseases. Among them, CXCL8 (interleukin-8, IL-8), binds to both the CXCR1 and CXCR2 receptors with high affinity and the expression levels of CXCL8 are elevated in many inflammatory diseases. Recently, an analogue of human CXCL8...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2008.04.007

    authors: Cheng HT,Huang KC,Yu HY,Gao KJ,Zhao X,Li F,Town J,Gordon JR,Cheng JW

    更新日期:2008-09-01 00:00:00

  • Purification and characterization of the hexokinase from Schistosoma mansoni, expressed in Escherichia coli.

    abstract::The hexokinase (ATP;D-hexose 6-phosphotransferase, EC 2.7.1.1) of Schistosoma mansoni has been expressed in Escherichia coli as a fusion protein including an N-terminal polyhistidine tag and enterokinase cleavage site. The enzyme was purified by metal chelate chromatography to > 95% homogeneity, based on analysis by S...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1996.0113

    authors: Armstrong RL,Wilson JE,Shoemaker CB

    更新日期:1996-11-01 00:00:00

  • Cloning, expression and functional characterization of two sesquiterpene synthase genes from moso bamboo (Phyllostachys edulis).

    abstract::The purpose of this work was to characterize the functions of two sesquiterpene synthase genes from moso bamboo (Phyllostachys edulis). Two novel sesquiterpene synthase genes, belonging to the Tpsa subfamily, were isolated from moso bamboo. MoTPS2 was 1641 bp in length and encoded a protein of 63 kDa, whereas MoTPS6 w...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2015.11.019

    authors: Chen X,Wang Y,Sun J,Wang J,Xun H,Tang F

    更新日期:2016-04-01 00:00:00

  • Isolation, purification and characterization of a novel solvent stable lipase from Pseudomonas reinekei.

    abstract::The Pseudomonas sp. have been long recognized for their exogenous lipolytic activities yet the genus still contains a lot of unexplored strains. Due to the versatile metabolic machinery and their potential for adaptation to fluctuating environmental conditions Pseudomonas sp. are of great interest for biotechnological...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2018.08.007

    authors: Priyanka P,Kinsella G,Henehan GT,Ryan BJ

    更新日期:2019-01-01 00:00:00

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

    abstract::The hemolytic phospholipase C/sphingomyelinase PlcH from the opportunistic pathogen Pseudomonas aeruginosa represents the founding member of a growing family of virulence factors identified in a wide range of bacterial and fungal pathogens. In P. aeruginosa PlcH is co-expressed with a 17 kDa chaperone (PlcR2) and secr...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2012.11.005

    authors: Truan D,Vasil A,Stonehouse M,Vasil ML,Pohl E

    更新日期:2013-07-01 00:00:00

  • Expression and one-step purification of intracellular human prolactin in insect cells.

    abstract::Human prolactin was expressed in insect culture cells by recombinant baculoviruses carrying prolactin gene cDNA placed under the transcriptional control of polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus. Preliminary results of recombinant human prolactin expression as extracellular as we...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.2001.1419

    authors: Strokovskaya L,Bartoszewicz Z,Szolajska E,Kikhno I,Solomko A,Michalik J

    更新日期:2001-07-01 00:00:00

  • The human rhabdomyosarcoma cell line TE671--Towards an innovative production platform for glycosylated biopharmaceuticals.

    abstract::The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-t...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2015.08.008

    authors: Rosenlöcher J,Weilandt C,Sandig G,Reinke SO,Blanchard V,Hinderlich S

    更新日期:2015-11-01 00:00:00

  • Production and characterization of biologically active human GM-CSF secreted by genetically modified plant cells.

    abstract::Human granulocyte-macrophage colony-stimulating factor (GM-CSF), a hemopoietic growth factor, was produced and secreted from tobacco cell suspensions. The GM-CSF cDNA was carried by a binary vector under the control of the CaMV 35S promoter and the T7 terminator. In addition, a 5'-nontranslated region from the tobacco...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.2000.1232

    authors: James EA,Wang C,Wang Z,Reeves R,Shin JH,Magnuson NS,Lee JM

    更新日期:2000-06-01 00:00:00

  • Purification, characterization, and crystallization of membrane bound Escherichia coli tyrosine kinase.

    abstract::Escherichia coli tyrosine kinase (Etk) is a membrane bound kinase in gram-negative bacteria that regulates the export of capsular polysaccharides (CPS). The molecular mechanism behind CPS regulation remains unclear, despite access to a crystal structure of the cytoplasmic kinase domain of Etk. In this study, an effici...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2015.08.029

    authors: Chesterman C,Jia Z

    更新日期:2016-09-01 00:00:00

  • Practical considerations in refolding proteins from inclusion bodies.

    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

    authors: Tsumoto K,Ejima D,Kumagai I,Arakawa T

    更新日期:2003-03-01 00:00:00

  • A combined approach for enhancing the stability of recombinant cis-dihydrodiol naphthalene dehydrogenase from Pseudomonas putida G7 allowed for the structural and kinetic characterization of the enzyme.

    abstract::The second enzyme of the naphthalene degradation pathway in Pseudomonas putida G7 is NahB, a dehydrogenase that converts cis-1,2-dihydroxy-1,2-dihydronaphthalene to 1,2-dihydroxynaphthalene. We report the cloning, optimization of expression, purification, kinetic studies and preliminary structural characterization of ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2017.01.005

    authors: Costa DMA,Costa MAF,Guimarães SL,Coitinho JB,Gómez SV,Brandão TADS,Nagem RAP

    更新日期:2017-04-01 00:00:00