Abstract：
:A synthetic gene encoding a single chain Fv fragment of an antibody directed against the nuclear inclusion a (NIa) protein of potato virus Y (PVY) was used to transform two commercial potato cultivars (Claustar and BF15). The NIa protease forms the nuclear inclusion body A and acts as the major protease in the cleavage of the viral polyprotein into functional proteins. Immunoblot analysis showed that most of the resulting transgenic plants accumulate high levels of the transgenic protein. Furthermore, a majority of the selected transgenic lines showed an efficient and complete protection against the challenge virus after mechanical inoculation with PVYO strain. Two transgenic lines showed an incomplete resistance with delayed appearance of symptoms accompanied by low virus titers, whereas one line developed symptoms during the first days after inoculation but recovered rapidly, leading to a low virus accumulation rate. These results confirm that expression of scFv antibody is able to inhibit a crucial step in the virus multiplication, such as polyprotein cleavage is a powerful strategy for engineered virus resistance. It can lead to a complete resistance that was not obtained previously by expression of scFv directed against the viral coat protein.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Gargouri-Bouzid R,Jaoua L,Rouis S,Saïdi MN,Bouaziz D,Ellouz Rdoi
10.1385/MB:33:2:133subject
Has Abstractpub_date
2006-06-01 00:00:00pages
133-40issue
2eissn
1073-6085issn
1559-0305pii
MB:33:2:133journal_volume
33pub_type
杂志文章abstract：:Enzymes are key molecules in signal-transduction pathways. However, only a small fraction of more than 500 human kinases, 300 human proteases and 200 human phosphatases is characterised so far. Peptide microarray based technologies for extremely efficient profiling of enzyme substrate specificity emerged in the last y...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/s12033-011-9402-x
更新日期：2011-11-01 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：:The Embrapa 5.1 genetically modified (GM) common bean was approved for commercialization in Brazil. Methods for the quantification of this new genetically modified organism (GMO) are necessary. The development of a suitable endogenous reference is essential for GMO quantification by real-time PCR. Based on this, a new...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9786-5
更新日期：2014-11-01 00:00:00
abstract：:Modern sugarcane cultivars have complex genetic characteristics and low fertility that render their genetic improvement through traditional breeding difficult. Genetic engineering methodology to introduce foreign genes provides new opportunities for the genetic improvement of sugarcane cultivars. One of prerequisites ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:28:2:113
更新日期：2004-10-01 00:00:00
abstract：:Lactococcus lactis is industrially important microorganism used in many dairy fermentations. Numerous genes and gene expression signals from this organism have now been identified and characterized. Recently, several naturally occurring, inducible gene-expression systems have also been described in L. lactis. The main...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/BF02760814
更新日期：1998-04-01 00:00:00
abstract：:Monoclonal antibodies (mAbs) are the fastest growing class of biopharmaceuticals reflecting their diverse applications in research and the clinic. The correct glycosylation of mAbs is required to elicit effector functions such as complement-dependent and antibody-dependent cell-mediated cytotoxicity, although these ma...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-012-9531-x
更新日期：2013-03-01 00:00:00
abstract：:Small ubiquitin-related modifier (SUMO) fusion system has been shown to be efficient for enhancing expression and preventing degradation of the target protein. We showed herein that SUMO fusion to human keratinocyte growth factor 2 (hKGF-2) gene was feasible and it significantly enhanced protein expression and its eff...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-008-9135-7
更新日期：2009-05-01 00:00:00
abstract：:The induction of several secondary metabolites in plants is one of the most commonly observed effects after the external addition of methyl jasmonate (MeJA). After the elicitation of Catharanthus roseus hairy roots with different concentrations of MeJA, changes in the accumulation of alkaloids such as ajmalicine, serp...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-008-9111-2
更新日期：2009-03-01 00:00:00
abstract：:Amplification of DNA or RNA sequences using the polymerase chain reaction (PCR) or reverse transcriptase PCR (RT-PCR) requires primers of an appropriate length to be designed. Two hexamer restriction primers, denoted as E101 and H301, which correspond to sequences of EcoRI and HindIII recognition sites, respectively, ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:14:1:01
更新日期：2000-01-01 00:00:00
abstract：:Alternaria blight caused by Alternaria brassicicola (Schwein.) Wiltshire and A. brassicae (Berk.) Sacc., is one of the most important disease of rapeseed-mustard, characterized by the formation of spots on leaves, stem, and siliquae with premature defoliation and stunting of growth. These two species are very difficul...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9784-7
更新日期：2014-11-01 00:00:00
abstract：:In this review I briefly describe the technique of protein engineering and indicate how the present state of knowledge allows proteins to be mutated to increase or decrease stability. I discuss experiments on both model proteins and those of relevance to the food industry and show how hydrophobic forces are a major dr...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/BF02921609
更新日期：1995-10-01 00:00:00
abstract：:Podophyllotoxin (PPT) is a plant natural product that serves as a precursor for the synthesis of many well-known chemotherapeutic drugs. The limited availability and high demand for the source plants of PPT have led to the exploration of alternative sources for this compound. In this study, we utilized the endophytic ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-015-9888-8
更新日期：2015-12-01 00:00:00
abstract：:While conducting our research on mutations in the human blood platelet glycoprotein Ib-alpha (GPIbalpha) gene, we detected an unusual deletion of 84 bp. This deletion took place in vitro, during PCR and between two direct repeats. It was observed that the deletion could be detected either by the direct sequencing of t...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-008-9059-2
更新日期：2008-09-01 00:00:00
abstract：:Many preclinical studies have shown RNA interference (RNAi) as a new promising way to treat various human diseases including cancer and virus infection and there is an increasing demand for the large-scale preparation of short interfering RNAs (siRNAs) at low cost. Data are accumulating to show that endoribonuclease-p...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:31:3:203
更新日期：2005-11-01 00:00:00
abstract：:The monomeric bacteriophage RNA polymerases allow the synthesis of virtually any RNA molecule in unlimited quantity. In this protocol, we describe the preparation of plasmid and PCR-derived templates. A basic transcription protocol is provided with several optional modifications. The use of RNA probes in Northern blot...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02761751
更新日期：1997-04-01 00:00:00
abstract：:A gene encoding maltogenic amylase from acidic Bacillus sp. US149 (maUS149) was cloned, sequenced and over-expressed in Escherichia coli. The nucleotide sequence analysis revealed an open reading frame (ORF) of 1749 bp encoding a protein of 582 residues. The alignment of deduced amino acid sequence revealed a relative...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-007-9017-4
更新日期：2008-03-01 00:00:00
abstract：:The significance of actin cytoskeleton on cell growth was historically studied using toxic drugs, such as cytochalasin. However, it is possible that unpredictable effects of these agents may have influenced the reported observations. In our study, we have established a drug-free system using cofilin overexpression to ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:31:1:001
更新日期：2005-09-01 00:00:00
abstract：:The biosensor field has grown enormously since the first demonstration of the biosensor concept by Leland C. Clark, Jr. in 1962. Today's biosensor market is dominated by glucose biosensors, mass-produced enzyme electrodes for the rapid self-diagnosis of blood glucose levels by diabetes sufferers. Here we take a histor...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:32:3:249
更新日期：2006-03-01 00:00:00
abstract：:D-Glucosamine is a commonly used dietary supplement that promotes cartilage health in humans. Metabolic flux analysis showed that D-glucosamine production could be increased by blocking three pathways involved in the consumption of glucosamine-6-phosphate and acetylglucosamine-6-phosphate. By homologous single-exchang...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-020-00257-9
更新日期：2020-08-01 00:00:00
abstract：:A bacterial artificial chromosome library of the causal agent of the Black Sigatoka leaf spot disease of banana and plantain, Mycosphaerella fijiensis, has been constructed using a non-sphaeroplasting technique and characterized using both homologous and heterologous probes. After first and a second size selection of ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-007-0016-2
更新日期：2007-05-01 00:00:00
abstract：:Human immunodeficiency virus types 1 and 2 (HIV-1, -2), the etiological agents of AIDS, are retroviruses that replicate in CD4+ T lymphocytes, monocytes, and macrophages. Early anti-HIV therapies were directed at the step in the virus life cycle that was considered the most readily blocked: the transcription of viral ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02821510
更新日期：1994-02-01 00:00:00
abstract：:Laccases are polyphenol oxidoreductases used in a number of industrial applications. Due to the increasing demand for these "green catalysis" enzymes, the identification and biochemical characterisation of their novel properties is essential. In our study, cloned Madurella mycetomatis laccase (mmlac) genes were hetero...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-020-00281-9
更新日期：2021-01-01 00:00:00
abstract：:In an effort to develop microalgae as a robust system for the production of valuable proteins, we analyzed some of the factors affecting recombinant protein expression in the chloroplast of the green alga Chlamydomonas reinhardtii. We monitored mRNA accumulation, protein synthesis, and protein turnover for three codon...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-010-9348-4
更新日期：2011-05-01 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：: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：:A duplex real-time PCR assay with melting curve analysis, using the EvaGreen fluorescence dye, was developed for rapid and reliable identification of bovine and caprine in ruminant feeds. The method merges the use of bovine (Bos taurus) and caprine (Capra hircus) specific primers that amplify small fragments (bovine 9...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9756-y
更新日期：2014-08-01 00:00:00
abstract：:This review outlines the various methods available for submitting sequence data to the EMBL Nucleotide Sequence Database. Depending on the type of sequence data and the facilities available to the submitter, one method may be more suitable than another. Recent developments have been the World Wide Web submission tool,...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/BF02762322
更新日期：1996-08-01 00:00:00
abstract：:Over the last decade transgenic mouse models have become a common experimental tool for unraveling gene function. During this time there has been a growing expectation that transgenes resemble the in vivo state as much as possible. To this end, a preference away from heterologous promoters has emerged, and transgene c...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:29:2:153
更新日期：2005-02-01 00:00:00
abstract：:A short (43-bp) A/T-rich stretch of DNA located in the intergenic region between the baiA2 and baiF genes from Eubacterium sp. strain VPI 12708 was amplified by polymerase chain reaction (PCR) and inserted in front of the Shine-Dalgarno (SD) sequences of three inefficiently-expressed Eubacterium sp. strain VPI 12708 g...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02789174
更新日期：1999-02-01 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
