Abstract：
:The gene that was inferred to encode the TyrR protein of Haemophilus influenzae Rd was synthesized by polymerase chain reaction and inserted into a T7-based expression vector. Methods were developed to overexpress the TyrR protein of H. influenzae in Escherichia coli and to purify the protein on a large scale. Both in vitro and in vivo functional comparisons of the H. influenzae and E. coli TyrR proteins were carried out. The TyrR protein of H. influenzae was able to bind in vitro to an operator target upstream of the aroF-tyrA gene of E. coli. In the presence of [gamma-S]ATP, the DNA binding ability of the H. influenzae TyrR protein was drastically reduced. Despite the much shorter peptide chain length (318 amino acid residues vs 513), the TyrR protein of H. influenzae was as active in repressing the aroF promoter as the TyrR protein of E. coli. Repression by both proteins was enhanced in the presence of tyrosine; however, the transcriptional activation function associated with the TyrR protein of E. coli could not be detected when the H. influenzae TyrR protein was expressed in E. coli. By computer analysis, at least five operator targets for TyrR were identified within the genomic DNA of H. influenzae. These observations show that the assignment of function to the tyrR gene of H. influenzae was correctly made. Further studies of the H. influenzae TyrR protein in comparison to its E. coli counterpart should provide valuable mechanistic information on transcriptional regulation in this system.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Zhu Q,Zhao S,Somerville RLdoi
10.1006/prep.1997.0757subject
Has Abstractpub_date
1997-07-01 00:00:00pages
237-46issue
2eissn
1046-5928issn
1096-0279pii
S1046-5928(97)90757-9journal_volume
10pub_type
杂志文章abstract：:To study the subunit structure and the active site of human immunodeficiency virus reverse transcriptase (RT), the enzyme was expressed in E. coli and purified to homogeneity in large quantities. The recombinant enzyme consists of two major polypeptides of 66,000 and 53,000 Da in equimolar amounts and a minor species ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/1046-5928(92)90005-h
更新日期：1992-08-01 00:00:00
abstract：:In this work we communicate the heterologous expression of a laccase from Coriolopsis gallica in Pichia pastoris. This enzyme has been reported to efficiently degrade a variety of pollutants such as industrial dyes. The expression strategy included using a previously reported modified α-factor preproleader for enhance...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2017.06.001
更新日期：2017-08-01 00:00:00
abstract：:A strategy for the purification and cleavage of chimeric recombinant proteins based on a genetically engineered metal-binding peptide and a human renin cleavage site is described. Vectors were constructed to direct the synthesis of chimeric human immunodeficiency virus (HIV) reverse transcriptase (RT) or beta-galactos...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/1046-5928(91)90073-r
更新日期：1991-04-01 00:00:00
abstract：:The B-subunits of replicative DNA polymerases belong to the superfamily of calcineurin-like phosphoesterases and are conserved from Archaea to humans. Recently we and others have shown that the B-subunit (DP1) of the archaeal family D DNA polymerase is responsible for proofreading 3'-5' exonuclease activity. The simil...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.05.002
更新日期：2005-09-01 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
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
更新日期：1999-07-01 00:00:00
abstract：:Plants possess very large numbers of biosynthetic cytochrome P450 enzymes. In spite of the importance of these enzymes for the synthesis of bioactive plant secondary metabolites, only two plant P450 structures has been obtained to date. Isoflavone synthase (IFS) is a membrane-associated cytochrome P450 enzyme catalyzi...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2010.11.012
更新日期：2010-12-04 00:00:00
abstract：:A tremendous increase has taken place over the last decades in the biochemical and clinical use of antibodies. Unfortunately, the constantly growing demand has not been matched by a corresponding easy access to pure immunoglobulin, as purification procedures tend to be either laborious, expensive, or inefficient. We p...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(03)00130-x
更新日期：2003-08-01 00:00:00
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
更新日期：2016-11-01 00:00:00
abstract：:Co-expression offers an important strategy for producing multiprotein complexes for biochemical and biophysical studies. We have found that co-expression of histones H2A and H2B (from yeast, chicken or Drosophila) leads to production of soluble heterodimeric H2AH2B complexes. Drosophila histones H3 and H4 can also be ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2010.03.013
更新日期：2010-08-01 00:00:00
abstract：:Streptococcal protein G has been used extensively for the purification of antibodies using the interaction of the Fc region with protein G. Many antibodies also interact with protein G through a low-affinity binding site for the Fab region. The exploitation of this low-affinity interaction for the purification of Fab'...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(05)80037-3
更新日期：1992-10-01 00:00:00
abstract：:Mitochondria-mediated apoptosis (programmed cell death) involves a sophisticated signaling and regulatory network that is regulated by the Bcl-2 protein family. Members of this family have either pro- or anti-apoptotic functions. An important pro-apoptotic member of this family is the cytosolic Bax. This protein is cr...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.02.004
更新日期：2019-06-01 00:00:00
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
更新日期：2008-10-01 00:00:00
abstract：:The gene from Aeromonas veronii bv. sobria encoding the metallo-beta-lactamase ImiS was subcloned into pET-26b, and ImiS was over-expressed in BL21(DE3) Escherichia coli and purified using SP-Sepharose chromatography. This protocol yielded over 5 mg of ImiS per liter of growth culture under optimum conditions. The bio...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.04.017
更新日期：2004-08-01 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：:Stromal-cell-derived factor-1 (SDF-1alpha) is an 8-kDa chemokine that is constitutively expressed in bone-marrow-derived stromal cells and has been identified as a ligand for the CXCR4 receptor. We produced the chemokine recombinantly as methionine-SDF-1alpha in Escherichia coli without the leader peptide sequence. Th...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2001.1402
更新日期：2001-04-01 00:00:00
abstract：:Ciliary neurotrophic factor (CNTF) is characterized as a neuropoietic cytokine for a broad spectrum of neurons, leading to its evaluation in humans suffering from neurodegenerative diseases. Due to its wide range of biological applications, high yield production of soluble biologically active recombinant human CNTF (r...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.01.008
更新日期：2014-04-01 00:00:00
abstract：:High-level constitutive expression of xylanase GH11 from Penicillium occitanis Pol6 termed PoXyn2 was achieved using the methylotrophic yeast Pichia pastoris. The PoXyn2 cDNA encoding for a mature xylanase of 320 amino acids was subcloned into the pGAPZαA vector, to construct recombinant xylanse with six histidine res...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.02.012
更新日期：2012-05-01 00:00:00
abstract：:A purification scheme based on hydrophobic interaction chromatography was developed to separate inactivated foot-and-mouth disease virus (FMDV) from crude supernatant. About 92% recovery and 8.8-fold purification were achieved on Butyl Sepharose 4 FF. Further purification on Superdex 200 resulted in another 29-fold pu...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.04.011
更新日期：2015-09-01 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
pub_type： 杂志文章
doi：10.1006/prep.1993.1051
更新日期：1993-10-01 00:00:00
abstract：:Glucose-6-phosphate dehydrogenase was purified from human placenta using DEAE-Sepharose fast flow, 2',5'-ADP Sepharose 4B column chromatography, and chromatofocusing on PBE 94 with PB 74. The enzyme was purified with 62% yield and had a specific activity of 87 units per milligram protein. The pH optimum was determined...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1370
更新日期：2001-03-01 00:00:00
abstract：:Proteus mirabilis (P. mirabilis) is a zoonotic pathogen that has recently presented a rising infection rate in the poultry industry. To develop an effective vaccine to protect chickens against P. mirabilis infection, OmpA, one of the major outer membrane proteins of P. mirabilis, was expressed in Pichia pastoris. The ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.10.001
更新日期：2015-01-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2009.05.011
更新日期：2009-11-01 00:00:00
abstract：:The iron-sulfur protein subunit, known as the Rieske protein, is one of the central components of the cytochrome b(6)f complex residing in chloroplast and cyanobacterial thylakoid membranes. We have constructed plasmids for overexpression in Escherichia coli of full-length and truncated Rieske (PetC) proteins from the...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(03)00016-0
更新日期：2003-05-01 00:00:00
abstract：:A method was developed for the affinity purification of human complement properdin. The preparation is part of an integrated scheme in which over 20 human plasma proteins can be recovered in a highly purified form. The yield of properdin was 5.9 mg from 3 liters of plasma, amounting to a 28% recovery. The crucial step...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1994.1026
更新日期：1994-04-01 00:00:00
abstract：:Human endothelin-1 (ET-1) is a potent vasocontractile 21-residue peptide hormone with significant pharmacological importance. An efficient and straightforward expression strategy that enables cost-effective incorporation of stable isotopes is not available thus far. In this report, we describe a cost-effective express...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.01.022
更新日期：2006-08-01 00:00:00
abstract：:The guanine nucleotide exchange factor EF-1beta gene from the thermoacidophilic archaeon Sulfolobus solfataricus (SsEF-1beta) was amplified by PCR and cloned into the pT7-7 expression vector. One of four selected clones harbored the T160C nucleotide substitution leading to the Y54H amino acid change in a hydrophobic r...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1997.0806
更新日期：1998-02-01 00:00:00
abstract：:The recently classified group III trypsins include members like Atlantic cod (Gadus morhua) trypsin Y as well as seven analogues from other cold-adapted fish species. The eight group III trypsins have been characterized from their cDNAs and deduced amino acid sequences but none of the enzymes have been isolated from t...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.06.008
更新日期：2007-02-01 00:00:00
abstract：:Lectins are a class of proteins with specific carbohydrate-binding properties found in a wide variety of plants and animals. Gramineae lectins are presumably defense-related proteins in plants that exert their effect by binding to N-acetylglucosamine. Barley lectin is a vacuolar protein synthesized with an amino-termi...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/1046-5928(92)90068-8
更新日期：1992-12-01 00:00:00
abstract：:Human cystatin C (HCC), encoded by cystatin 3 gene, is a 13.3kDa endogenous cysteine proteinase inhibitor and an important biomarker of renal function. However, expressing recombinant cystatin C is difficult because of low yield and inclusion bodies in Escherichia coli (E. coli). In this study, we cloned HCC gene into...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.03.006
更新日期：2015-07-01 00:00:00
