Abstract:
:The filamentous fungus Sclerotinia sclerotiorum produces a complete set of cellulolytic enzymes needed for efficient solubilization of native cellulose, the major component of plants. In this work, we reported the molecular characterization of an important glycosyl-hydrolase enzyme classified as endo-β-1,4-glucanase. The importance of this enzyme was revealed with the in-gel activity staining, showing a high degradation capacity of cellulose. When purified from native gel and ran in denaturing polyacrylamide gel, the polypeptide has an apparent molecular mass of about 34 kDa called Endo2. For further characterization of this protein, a mass spectrometry approach was carried out. The LC-MS/MS analysis revealed two peptides belonging to this enzyme. The genomic DNA and cDNA sequences were resolved by PCR amplification and sequencing, revealing a gene with two intron sequences. The open reading frame of 987 bp encoded a putative polypeptide of 328 amino acids having a calculated molecular mass of 33,297 Da. Yet, the molecular modeling and comparative investigation of different 3D cellulase structures showed that this endoglucanase isoform has probably two domains. A core domain having a high similarity with endoglucanases family 5 and a cellulose-binding domain having similarities with those of exo-type cellulases of family 1, linked together by a serine-threonine-rich region. These results are with great interests and show new characteristics of S. sclerotiorum glucanase.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Chahed H,Ezzine A,Mlouka AB,Hardouin J,Jouenne T,Marzouki MNdoi
10.1007/s12033-013-9714-0subject
Has Abstractpub_date
2014-04-01 00:00:00pages
340-50issue
4eissn
1073-6085issn
1559-0305journal_volume
56pub_type
杂志文章abstract::Considering the fact that Prays oleae is one of the most pathogenic insects to the olive tree in the Mediterranean basin, particularly in Tunisia, the mode of action of Cry insecticidal toxins of Bacillus thuringiensis kurstaki in Prays oleae midgut was investigated. The proteolysis of Bacillus thuringiensis delta-end...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02686109
更新日期:2007-02-01 00:00:00
abstract::In order to further study the B subunit of the Escherichia coli heat-labile enterotoxin (LTB), we obtained the LTB gene from pathogenic E. coli, cloned it into the pET22b (+) prokaryotic expression vector, and expressed it as a fusion protein with His tag in E. coli BL21 (DE3). The recombinant LTB was expressed and pu...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9101-4
更新日期:2009-02-01 00:00:00
abstract::This article describes the construction, expression, and purification of RNase single-chain antibody fusion proteins. To construct a fusion protein, the gene for each moiety, the RNase and the binding ligand, is modified separately to contain complementary DNA encoding a 13 amino acid spacer that separates the RNase f...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1385/MB:20:1:063
更新日期:2002-01-01 00:00:00
abstract::G-quartets are square planar arrangements of four guanine bases, which can form extraordinarily stable stacks when present in nucleic acid sequences. Such G-quadruplex structures were long regarded as an in vitro phenomenon, but the widespread presence of suitable sequences in genomes and the identification of protein...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s12033-011-9395-5
更新日期:2011-10-01 00:00:00
abstract::D-Stereospecific amidohydrolase (DAH) from Streptomyces sp. 82F2 has potential utility for the synthesis of D/L configuration dipeptides by an aminolysis reaction. Structural comparison of DAH with substrate-bound D-amino acid amidase revealed that three residues located in the active site pocket of DAH (Thr145, Ala26...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-018-0104-5
更新日期:2018-09-01 00:00:00
abstract::Biobanks, more formally known as biological resource centers (BRCs), form an "unsung" yet critical component of the infrastructures for scientific research, industry and conservation, without which much of the current scientific activity involving microbial cultures and cell-lines would be effectively impossible. BRCs...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s12033-008-9099-7
更新日期:2008-10-01 00:00:00
abstract::Enterotoxigenic strains of Escherichia coli produce a heat-labile holotoxin (LT), which causes diarrhea. We engineered corn seeds to produce LT-B, the nontoxic subunit of LT, to serve as a plant-derived vaccine to traveler's diarrhea and as an adjuvant for co-administered proteins. We previously demonstrated that a st...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:32:1:031
更新日期:2006-01-01 00:00:00
abstract::A sandwich-type enzyme-linked immunosorbent assay (ELISA) was established for monitoring the secretion of ZZ-fusion proteins. Two antibodies, a monoclonal mouse anti-human proinsulin and a rabbit anti-bovine IgG (strongly binding to the ZZ-domain), were used to quantify the secretion of recombinant human ZZ-proinsulin...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:19:3:239
更新日期:2001-11-01 00:00:00
abstract::In the present study, four full-length Dof (DNA-binding with one finger) genes from Sorghum bicolor namely SbDof1, SbDof19, SbDof23, and SbDof24 were PCR amplified, gel eluted, cloned, and sequenced (accession number HQ540084, HQ540085, HQ540086, and HQ540087, respectively). These sequences were further characterized ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9536-5
更新日期:2013-05-01 00:00:00
abstract::The enzyme avian myeloblastosis virus reverse transcriptase (AMV-RT) is routinely used for cDNA synthesis, which is generally carried out at temperatures between 37 degrees C and 42 degrees C. We show that this enzyme can support cDNA synthesis, at temperatures as high as 70 degrees C. We have utilized this property o...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:12:3:237
更新日期:1999-10-01 00:00:00
abstract::The recombinant acidstable α-amylase (Ba-amy) of acidophilic bacterium Bacillus acidicola TSAS1 has been produced extracellularly using a combination of cloning (E. coli and P. pastoris) and physico-chemical treatment strategies. A total of 150,000 U/L of Ba-amy were attained under constitutive promoter in P. pastoris...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-016-9970-x
更新日期:2016-11-01 00:00:00
abstract::Isolation of RNA from recalcitrant tree tissues has been problematic due to large amounts of secondary metabolites and interfering compounds in their cells. We have developed an efficient RNA extraction method, which yielded high-quality RNA preparations from tissues of the lychee tree. The method reported here utiliz...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-007-0073-6
更新日期:2008-01-01 00:00:00
abstract::Several mutations were introduced into the Cry1Ac toxin gene, resulting in four variants with altered sequences that were responsible for low expression of the toxin in transgenic plants. These variants were as follows: V1, with modified three A/T-rich regions, including the first signal of transcription termination; ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/mb:26:1:17
更新日期:2004-01-01 00:00:00
abstract::D-Allulose is a rare monosaccharide that exists in extremely small quantities in nature, and it is also hard to prepare at a large scale via chemical or enzyme synthetic route due to low conversion and downstream separation complexity. Using D-psicose epimerase and L-rhamnulose kinase, a method enabling high conversio...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-019-00174-6
更新日期:2019-06-01 00:00:00
abstract::Group B Streptococcus (GBS) is the leading cause of neonatal meningitis and a common pathogen in livestock and aquaculture industries around the world. Conjugate polysaccharide and protein-based vaccines are under development. The surface immunogenic protein (SIP) is a conserved protein in all GBS serotypes and has be...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-018-0065-8
更新日期:2018-03-01 00:00:00
abstract::Production of monoclonal antibodies and pharmaceutical proteins in transgenic plants has been the focus of many research efforts for close to 30 years. Use of plants as bioreactors reduces large-scale production costs and minimizes risk for human pathogens contamination. Stable nuclear transformation of the plant geno...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-018-0075-6
更新日期:2018-06-01 00:00:00
abstract::Megaprimer-based methodology has been widely applied in site-directed mutagenesis, but rarely used in gene splicing. In this article, we describe a modification of the megaprimer PCR method, which can efficiently create and amplify a specific ligated chimeric gene segment in a PCR reaction and under a common PCR progr...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9078-z
更新日期:2008-11-01 00:00:00
abstract::Adeno-associated virus (AAV) is a small, non-enveloped virus used as vector in gene therapy, mainly produced in human cells and in baculovirus systems. Intense studies on these platforms led to the production of vectors with titers between 103 and 105 viral genomes (vg) per cells. In spite of this, vector yields need ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-020-00294-4
更新日期:2021-02-01 00:00:00
abstract::Papaya ringspot virus (PRSV) causes severe economic losses in both cucurbits and papaya throughout the tropics and subtropics. Development of PRSV-resistant transgenic plants faces a major hurdle in achieving resistance against geographically distinct isolates. One of the major reasons of failing to achieve the broad-...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-009-9205-5
更新日期:2010-01-01 00:00:00
abstract::Due to the limited number of molecular studies focused on European gene pool investigation, it is necessary to perform plant material recognition. Eighteen accessions of three Miscanthus species, namely, M. × giganteus, M. sinensis, M. sacchariflorus were evaluated with the use of molecular marker systems such as: int...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-014-9770-0
更新日期:2014-10-01 00:00:00
abstract::Recombinant adenovirus vectors have proven to be useful tools in facilitating gene transfer. Construction of such vectors requires a knowledge of the adenovirus genome structure and its life cycle. A commonly used recombinant adenovirus involves deletion of the E1 region; such a recombinant is traditionally produced b...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:18:1:63
更新日期:2001-05-01 00:00:00
abstract::Malaria is a serious but preventable and treatable infectious disease that is found in over 100 countries around the world. Correct and rapid diagnosis of malaria infection can rescue the patient of getting sicker and reduces the risk of disease spreading among humans. Chlamydomonas reinhardtii chloroplast is an attra...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-018-0140-1
更新日期:2019-02-01 00:00:00
abstract::In this study cationic β-cyclodextrin-chitosan-mediated nanoparticles were used to transfer pmCherry-C1 into glioblastoma cells and their transfection efficiency were compared to lipofectamine and electroporation. Physicochemical characteristics of nanoparticles were evaluated by photon correlation spectroscopy and sc...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-016-9927-0
更新日期:2016-04-01 00:00:00
abstract::DNA beta is an approx 1350 nucleotide, single-stranded DNA molecule which has been shown to be associated with some monopartite geminiviruses of the genus Begomovirus. This component requires the helper begomovirus for replication in the cells of host plants and for insect transmission, possibly by trans-encapsidation...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:20:3:315
更新日期:2002-03-01 00:00:00
abstract::This article presents a simple and rapid method for removal of unincorporated label and proteins from DNA sequencing reactions by using Wizard purification resin. This method can be successfully applied for preparation of end-labeled oligonucleotides free of unincorporated label, which is important in experiments (inc...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02789066
更新日期:1996-04-01 00:00:00
abstract::Molecular biological methods that use antibodies and nucleic acids to detect specific foodborne bacterial pathogens were scarcely known a decade and a half ago. Few scientists could have predicted that these tools of basic research would come to dominate the field of food diagnostics. Today, a large number of cleverly...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/BF02740817
更新日期:1997-06-01 00:00:00
abstract::Dye-based protein determination assays are widely used to estimate protein concentration, however various reports suggest that the response is dependent on the composition and sequence of the protein, limiting confidence in the resulting concentration estimates. In this study a diverse set of model proteins representi...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-007-0038-9
更新日期:2007-10-01 00:00:00
abstract::The MYB proteins constitute one of the largest transcription factor families in plants. Much research has been performed to determine their structures, functions, and evolution, especially in the model plants, Arabidopsis, and rice. However, this transcription factor family has been much less studied in wheat (Triticu...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-011-9486-3
更新日期:2012-10-01 00:00:00
abstract::Angiogenesis has been considered an important target for cancer therapy. The inhibition of angiogenesis represents a promising strategy for anti-cancer treatment, tumor growth inhibition, and metastasis. Vasostatin 30 (Vs30), and the 14.1 kDa vasoinhibin (Vi-II-14.1) are two peptides with remarkable anti-tumor and ant...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9642-4
更新日期:2013-07-01 00:00:00
abstract::Agrobacterium-mediated transformation is the method of choice to engineer desirable genes into plants. Here we describe a protocol for demonstrating T-DNA transfer from Agrobacterium into the economically important graminaceous plant maize. Expression of the T-DNA-located GUS gene was observed with high efficiency on ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:13:2:165
更新日期:1999-12-01 00:00:00