Abstract:
:We report on a unified rapid betaine-based-PCR protocol for amplification of the (CAG)n region in Huntington disease (HD) and the (CGG)n region in Fragile X syndrome (FXS), followed by an electrophoretic separation on automated sequencer for precise determination of the triplet numbers. The high betaine concentration (2.5 M betaine) permits precise amplification of the CAG and CGG repeats. Ten HD affected patients and 10 healthy individuals from HD families were re-evaluated. For FXS the CGG region in normal individuals and premutations of about 100 repeats were precisely amplified by this protocol. Ten unrelated FXS premutation carriers and 24 mentally retarded non-FXS affected boys were re-examined by this method. The results totally coincided with the previous ones. This protocol is a good choice as a fast screening test. Within 24 h we can have preliminary information on the patient's genetic status. Normal individuals, CGG premutation carriers up to 100 repeats, as well as HD patients carrying an expansion up to 50 CAG repeats can be easily clarified. This accounts for a relatively large proportion (about 90%) of the suspected HD and FXS patients, referred to our laboratory for genetic analysis. The calculation of the repeat's number is more accurate for the correct interpretation of the results, screening tests and genetic counselling.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Todorov T,Todorova A,Georgieva B,Mitev Vdoi
10.1007/s12033-010-9260-ysubject
Has Abstractpub_date
2010-06-01 00:00:00pages
150-4issue
2eissn
1073-6085issn
1559-0305journal_volume
45pub_type
杂志文章abstract::An alternative method to combine mutagenesis PCR with dITP and fragmentation by endonuclease V for directed evolution was developed. In comparison to the routine protocol for directed evolution, dITP was used as mutation reagent in the mutagenesis PCR. Subsequently, the incorporated dITP in the PCR products could repr...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9516-9
更新日期:2013-01-01 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::We describe an expression system for high-yield production of recombinant soluble human FasL (rsh- FasL) in CHO cells. After one round of selection for gene amplification, cell lines producing rsh-FasL up to 60 microg/L x 10(6) cells in 24 h were obtained. Cell lines were grown in protein-free medium as suspension cul...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:23:3:189
更新日期:2003-03-01 00:00:00
abstract::Efficient display of antibody on filamentous phage M13 coat is crucial for successful biopanning selections. We applied a directed evolution strategy to improve the oligovalent display of a poorly behaving Fab fragment fused to phage gene-3 for minor coat protein (g3p). The Fab displaying clones were enriched from a r...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-009-9231-3
更新日期:2010-03-01 00:00:00
abstract::Adipose tissue-derived mesenchymal stromal cells (ADSCs) are a prominent cellular source for regenerative medicine. We tested whether transplantation of ADSCs into the ischemic muscular tissue of diabetic animals would attenuate impaired cell metabolism and microcirculatory function. We induced unilateral hind limb is...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-014-9735-3
更新日期:2014-05-01 00:00:00
abstract::The ability to identify disease-associated genes using microarray technology is dependent on isolating high-quality total RNA from the diseased tissue under study. However, it is not always possible to obtain large amounts of affected tissue from patients and therefore the quality and yield of RNA may be compromised. ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:23:2:171
更新日期:2003-02-01 00:00:00
abstract::Water-soluble quinoprotein glucose dehydrogenase (PQQGDH-B) is a dimeric enzyme whose application for glucose sensing is the focus of much attention. We attempted to increase the thermal stability of PQQGDH-B by introducing a disulfide bond at the dimer interface. The Ser residue at position 415 was selected for subst...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:24:2:97
更新日期:2003-06-01 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::In mammals, CpG methylation is one of the mechanisms of epigenetic control over the linear sequence of bases of deoxyribonucleic acid (DNA); about 70% of CpG dinucleotides are methylated. The actual signal that triggers DNA methylation is not known, although repetitive DNA has been shown to be an attractive template f...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:26:3:215
更新日期:2004-03-01 00:00:00
abstract::It is difficult to obtain precise and accurate estimates of the true mycotoxin concentration of a bulk lot when using a mycotoxin-sampling plan that measures the concentration in a small portion of the bulk lot. A mycotoxin-sampling plan is defined by a mycotoxin test procedure and a defined accept/reject limit. A myc...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1385/MB:23:1:61
更新日期:2003-01-01 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::An endochitinase gene from the Serratia marcescens Nima strain (chiA Nima) was cloned, sequenced, and expressed in Escherichia coli DH5alphaF', and the recombinant protein (ChiA Nima) was purified by hydrophobic interaction chromatography. chiA Nima contains an open reading frame (ORF) that encodes an endochitinase wi...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:31:2:103
更新日期:2005-10-01 00:00:00
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::The clinical management of immunocompromised patients depends on the rapid identification of infectious agents such as fungal pathogens. The procedure described here for accomplishing this uses a sensitive polymerase chain reaction method, previously reported, combined with restriction-enzyme digestion to distinguish ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:24:3:229
更新日期:2003-07-01 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::Clinical molecular genetics has recently become recognized as a diagnostic discipline. This article covers the evolution, structure, and possible forward development of clinical molecular genetics. Topics covered include general test categories, introducing new tests, laboratory facilities, staffing and training, and ...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/BF02752254
更新日期:1997-10-01 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 polyamines spermine, spermidine, and putrescine are small organic molecules one or more of which are present in all living organisms. Many natural products contain polyamine residues. Polyamines are synthesized by a highly regulated pathway from arginine or ornithine and also can be transported in and out of cells...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/BF02788682
更新日期:1999-06-01 00:00:00
abstract::The concept of photoprotection by dietary means is gaining momentum. Plant constituents such as carotenoids and flavonoids are involved in protection against excess light in plants and contribute to the prevention of UV damage in humans. As micronutrients, they are ingested with the diet and are distributed into light...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s12033-007-0051-z
更新日期:2007-09-01 00:00:00
abstract::A naturally secreted Gaussia luciferase (Gluc) has been utilized as a reporter for bioluminescence imaging (BLI) evaluation. However, the potential application of Gluc for in vivo monitoring of systemic protein delivery, as well as its natural biodistribution, has not been studied. To examine Gluc secretion and uptake...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9519-6
更新日期:2013-01-01 00:00:00
abstract::RNA editing is one of the post-transcriptional RNA processes. RNA editing generates RNA and protein diversity in eukaryotes and results in specific amino acid substitutions, deletions, and changes in gene expression levels. Adenosine-to-inosine RNA editing represents the most important class of editing in human and af...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/s12033-012-9498-7
更新日期:2012-09-01 00:00:00
abstract::The REDOX-RESPONSIVE TRANSCRIPTION FACTOR 1 (RRTF1) gene encodes a member of the ERF/AP2 transcription factor family involved in redox homeostasis. The RRTF1 gene shows tissue-specific responsiveness to various abiotic stress treatments including a response to salt stress in roots. An interesting feature of this respo...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-019-00155-9
更新日期:2019-04-01 00:00:00
abstract::Artificial microRNA (amiRNA) has become a powerful tool for gene silencing in plants. A new method for easy and rapid construction of rice artificial miRNA vector is described. The procedure involved modification of the pCAMBIA1300-UR vector by insertion of a 'vector modification fragment'. This was prepared from the ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-010-9291-4
更新日期:2010-11-01 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::The macrofungus Cordyceps militaris contains many kinds of bioactive ingredients that are regulated by functional genes, but the functions of many genes in C. militaris are still unknown. In this study, to improve the frequency of homologous integration, a genetic transformation system based on a split-marker approach...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-018-0080-9
更新日期:2018-05-01 00:00:00
abstract::Previous studies have shown that mRNA-electroporated dendritic cells (DCs) are able to process and present tumor-associated antigens, leading to the activation of tumor-specific T cells in vitro and in vivo. However, the optimal maturation state of antigen loading and half-life of the mRNA-translated protein product a...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9071-6
更新日期:2008-10-01 00:00:00
abstract::Surface plasmon resonance (SPR) biosensor is a promising technology for its various advantages including the real-time measurement of biomolecular interactions without labeling. A method of hybridizing RNAs on the surface of the streptavidin-coated (SA) sensor chip to study RNA-protein interactions was described in th...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9066-3
更新日期:2008-09-01 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::We report a rapid and simple method for isolating the 5'-end of plant genes from genomic DNA by polymerase chain reaction (PCR) with TATA-box-based degenerate primers (TDPs). The TDPs were specially designed according to the TATA box, which is conserved in the promoter region of most plant genes. The unknown 5'-ends o...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-010-9323-0
更新日期:2011-02-01 00:00:00
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. ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-013-9714-0
更新日期:2014-04-01 00:00:00