Abstract:
:In this study, we investigated the relative expression of the Rous sarcoma virus (RSV) promoter-driven expression of enhanced green fluorescent protein (EGFP) in fibroblasts of transgenic quails. We analyzed the direct influence of CpG methylation of the RSV promoter on the transcriptional activity of delivered transgenes. Embryonic fibroblasts collected from homozygous transgenic quail (TQ2) were treated with 50 μM of DNA methyltransferase inhibitor followed by 5-aza-2'-deoxycytidine (5-azadC) for 48 h, and changes in expression were then analyzed by flow cytometry. The results show a significant increase of EGFP expression in TQ2 embryonic fibroblasts (QEFs) (2.64% to 79.84%). Subsequent methylation-specific amplification revealed that 5-azadC significantly reduced the CpG methylation status in the RSV promoters of the QEFs (86.42 to 48.41%); even after 5-azadC was withdrawn, CpG methylation remained decreased in expanded culture (16.28%). Further analysis showed that potential transcription factor binding sites existed in the CpG methylation site of the RSV promoter. These results may provide the basis for understanding the epigenetic mechanism responsible for transgenic animal production and genetic preservation.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Jang HJ,Choi JW,Kim YM,Shin SS,Lee K,Han JYdoi
10.1007/s12033-011-9393-7subject
Has Abstractpub_date
2011-11-01 00:00:00pages
222-8issue
3eissn
1073-6085issn
1559-0305journal_volume
49pub_type
杂志文章abstract::Positional cloning of rare disease genes depends on the availability of highly polymorphic markers near the disease loci. The most abundant class of polymorphic markers in the human genome is CA-repeats. We have developed a strategy for the rapid isolation of highly polymorphic CA-repeats from YAC clones. Total DNA of...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02789104
更新日期:1995-04-01 00:00:00
abstract::De novo design provides an in silico toolkit for the design of novel small molecular structures to a set of specified structural constraints. With the avalanche of bioinformatics data, de novo design is ideally suited for exploring molecules that could be useful for chemical genomics. The design process involves manip...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s12033-007-0037-x
更新日期:2007-11-01 00:00:00
abstract::The diversity and complexity of bioinformatics tools currently available for protein sequence analysis can make it difficult to know where to begin when presented with a new sequence. In this article, we present a protocol outlining one approach to sequence analysis that should give as comprehensive a picture as possi...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-007-0032-2
更新日期:2007-07-01 00:00:00
abstract::The aim of this study was to evaluate the usefulness of replicating but non disseminating adenovirus vectors (AdVs) as vaccine vector using human rotavirus (HRV) as a model pathogen. HRV VP7, VP4, or VP4Δ (N-terminal 336 amino acids of VP4) structural proteins as well as the VP4Δ::VP7 chimeric fusion protein were expr...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-013-9653-9
更新日期:2013-07-01 00:00:00
abstract::In this study, host-specific forms of the blast pathogen Magnaporthe oryzae in sub-Saharan Africa (SSA) were characterised from distinct cropping locations using a combination of molecular and biological assays. Finger millet blast populations in East Africa revealed a continuous genetic variation pattern and lack of ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-011-9429-z
更新日期:2012-02-01 00:00:00
abstract::Deficiency of the lysosomal glucocerebrosidase (GCR) enzyme results in Gaucher's disease, the most common inherited storage disorder. Treatment consists of enzyme replacement therapy by the administration of recombinant GCR produced in Chinese hamster ovary cells. The production of anti-GCR antibodies has already been...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-010-9303-4
更新日期:2010-11-01 00:00:00
abstract::The advent of recombinant antibody technology led to an enormous revival in the use of antibodies as diagnostic and therapeutic tools for fighting cancer. This review provides a brief historical sketch of the development of recombinant antibodies for the diagnosis and immunotherapy of cancer and summarizes the most si...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1385/MB:25:1:1
更新日期:2003-09-01 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::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::The synthesis of complete genes is becoming a more and more popular approach in heterologous gene expression. Reasons for this are the decreasing prices and the numerous advantages in comparison to classic molecular cloning methods. Two of these advantages are the possibility to adapt the codon usage to the host organ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-011-9479-2
更新日期:2012-10-01 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::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::Genetically engineered animal models have been and will continue to be invaluable for exploring the basic mechanisms involved in the aging process as well as in extending our understanding of diseases found to be more prevalent in the older human population. Continued development of such in vivo systems will allow sci...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1385/MB:19:1:045
更新日期:2001-09-01 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
abstract::IgG class antibodies express catalytic activities rarely and at very low levels. Here, we studied polyclonal IgA and IgG preparations from healthy human sera and saliva for the ability to hydrolyze model peptidyl-aminomethylcoumarin (peptide-AMC) substrates. These substrates permit objective evaluation of the catalyti...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-007-0003-7
更新日期:2007-06-01 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::Coptotermes curvignathus is a termite that, owing to its ability to digest living trees, serves as a gold mine for robust industrial enzymes. This unique characteristic reflects the presence of very efficient hydrolytic enzyme systems including cellulases. Transcriptomic analyses of the gut of C. curvignathus revealed...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-017-0015-x
更新日期:2017-07-01 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::Isolation of high quality nucleic acids from plant tissues rich in polysaccharides and polyphenols is often difficult. The presence of these substances can affect the quality and/or quantity of the nucleic acids isolated. Here, we describe a rapid and efficient nucleic acids extraction protocol that in contrast to oth...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-011-9384-8
更新日期:2011-10-01 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::RNA interference methodology suppresses specific gene expression, thus mimicking loss-of-function mutation and enabling in vitro and in vivo gene function analysis. Lipofectamine RNAiMAX, a new transfection reagent, has been confirmed high efficiency in delivering small interfering RNA (siRNA) into mesenchymal stem ce...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9043-x
更新日期:2008-09-01 00:00:00
abstract::The term "coincidence cloning" encompasses a wide range of methodologies, the aim of which is to isolate DNA sequences which occur in both of two input DNA sources. The nature of these input DNAs may be genomic or cDNA, cloned or uncloned, and as such the far reaching applicability of the techniques can be imagined. I...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/BF02900362
更新日期:1996-06-01 00:00:00
abstract::We have modified the differential display of 3'-end restriction fragments of cDNA technique by combining it with the amplified fragment length polymorphism (AFLP) approach and silver staining. Modified oligo-dT primers were used for a reverse transcription step. ds cDNA was digested with the Mse I restriction enzyme a...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:12:2:137
更新日期:1999-09-01 00:00:00
abstract::To develop combined vaccine systems, we have generated Edwardsiella tarda ghosts (ETG) displaying a foreign protein on the outer membrane and also Ed. tarda cadaver (ETC) expressing a heterologous protein in the cytoplasm. Green fluorescent protein (GFP) was used as a model foreign protein. A constitutive promoter (Et...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-010-9277-2
更新日期:2010-10-01 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::Translation initiation of Coxsackievirus B3 (CVB3) RNA is directed by an internal ribosome entry site (IRES) within the 5' untranslated region. Host cell factors involved in this process include some canonical translation factors and additional RNA-binding proteins. We have, previously, described that the Sabin3-like ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9592-x
更新日期:2013-06-01 00:00:00
abstract::Chlorella ellipsoidea is a single-celled eukaryotic green microalgae with high nutritional value. Its value may be further increased if a simple, reliable and cost-effective transformation method for C. ellipsoidea can be developed. In this paper, we describe a novel transformation method for C. ellipsoidea . This sys...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9554-3
更新日期:2013-06-01 00:00:00
abstract::Dendritic cell-based vaccines are being evaluated in clinical trials to determine their ability to activate clinically relevant tumor antigen-specific immune responses. Although some groups isolate dendritic cells from peripheral blood, most have found it more efficient to generate large numbers from peripheral blood ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:25:1:95
更新日期:2003-09-01 00:00:00
abstract::Neuroblastoma is the most common extracranial solid tumor of infancy. Amplification of MYCN oncogene is found in approximately 20 % of all neuroblastoma patients and correlates with advanced disease stages, rapid tumor progression, and poor prognosis, making this gene an obvious therapeutic target. However, being a tr...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-014-9739-z
更新日期:2014-07-01 00:00:00
abstract::Identification of expressed sequence tags (ESTs) in large genomic segments is an important step in positional cloning and genomic mapping studies. A simple and efficient polymerase chain reaction (PCR)-based approach is described here to identify coding sequences in large genomic fragments of DNA cloned in vectors suc...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02760779
更新日期:1997-12-01 00:00:00