A unified rapid PCR method for detection of normal and expanded trinucleotide alleles of CAG repeats in huntington chorea and CGG repeats in fragile X syndrome.

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 Biotechnol

journal_title

Molecular biotechnology

authors

Todorov T,Todorova A,Georgieva B,Mitev V

doi

10.1007/s12033-010-9260-y

subject

Has Abstract

pub_date

2010-06-01 00:00:00

pages

150-4

issue

2

eissn

1073-6085

issn

1559-0305

journal_volume

45

pub_type

杂志文章
  • A simple and reproducible method for directed evolution: combination of random mutation with dITP and DNA fragmentation with endonuclease V.

    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

    authors: Wang Z,Wang HY,Feng H

    更新日期:2013-01-01 00:00:00

  • Investigation and Expression of the Secoisolariciresinol Dehydrogenase Gene Involved in Podophyllotoxin Biosynthesis.

    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

    authors: Arneaud SL,Porter JR

    更新日期:2015-12-01 00:00:00

  • Expression and characterization of biologically active human Fas ligand produced in CHO cells.

    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

    authors: Zappitelli S,D'Alatri L,Ciucci A,Raucci G,Faiella A,Gabrielli M,Parlani M,Bressan A,Maggi CA,Goso C,Rotondaro L

    更新日期:2003-03-01 00:00:00

  • Oligovalent Fab display on M13 phage improved by directed evolution.

    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

    authors: Huovinen T,Sanmark H,Ylä-Pelto J,Vehniäinen M,Lamminmäki U

    更新日期:2010-03-01 00:00:00

  • Transplantation of mesenchymal cells improves peripheral limb ischemia in diabetic rats.

    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

    authors: Madonna R,Delli Pizzi S,Tartaro A,De Caterina R

    更新日期:2014-05-01 00:00:00

  • Improved method for generating microarray probes using submicrogram amounts of total RNA.

    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

    authors: Murray EJ

    更新日期:2003-02-01 00:00:00

  • Stabilization of quaternary structure of water-soluble quinoprotein glucose dehydrogenase.

    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

    authors: Igarashi S,Sode K

    更新日期:2003-06-01 00:00:00

  • Miscanthus: genetic diversity and genotype identification using ISSR and RAPD markers.

    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

    authors: Cichorz S,Gośka M,Litwiniec A

    更新日期:2014-10-01 00:00:00

  • Transgene methylation in mice reflects copy number but not expression level.

    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

    authors: Pena RN,Webster J,Kwan S,Korbel J,Whitelaw BA

    更新日期:2004-03-01 00:00:00

  • Detecting mycotoxins in agricultural commodities.

    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

    authors: Whitaker TB

    更新日期:2003-01-01 00:00:00

  • The Optimisation of the Expression of Recombinant Surface Immunogenic Protein of Group B Streptococcus in Escherichia coli by Response Surface Methodology Improves Humoral Immunity.

    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

    authors: Díaz-Dinamarca DA,Jerias JI,Soto DA,Soto JA,Díaz NV,Leyton YY,Villegas RA,Kalergis AM,Vásquez AE

    更新日期:2018-03-01 00:00:00

  • Molecular cloning and purification of an endochitinase from Serratia marcescens (Nima).

    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

    authors: Ruiz-Sanchez A,Cruz-Camarillo R,Salcedo-Hernandez R,Ibarra JE,Barboza-Corona JE

    更新日期:2005-10-01 00:00:00

  • Rapid identification of polymorphic CA-repeats in YAC clones.

    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

    authors: Rotman G,Vanagaite L,Collins FS,Shiloh Y

    更新日期:1995-04-01 00:00:00

  • A simple PCR/RFLP analysis can differentiate between Candida albicans, Aspergillus niger, and Aspergillus fumigatus.

    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

    authors: Isik N,White L,Barnes R,Poynton CJ,Mills KI

    更新日期:2003-07-01 00:00:00

  • Biobanking.

    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

    authors: Day JG,Stacey GN

    更新日期:2008-10-01 00:00:00

  • An overview of clinical molecular genetics.

    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

    authors: Elles R

    更新日期:1997-10-01 00:00:00

  • Classification of protein folds.

    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

    authors: Russell RB

    更新日期:2007-07-01 00:00:00

  • Polyamines. An overview.

    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

    authors: Morgan DM

    更新日期:1999-06-01 00:00:00

  • Carotenoids and flavonoids contribute to nutritional protection against skin damage from sunlight.

    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

    authors: Stahl W,Sies H

    更新日期:2007-09-01 00:00:00

  • Secreted luciferase for in vivo evaluation of systemic protein delivery in mice.

    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

    authors: El-Amouri SS,Cao P,Miao C,Pan D

    更新日期:2013-01-01 00:00:00

  • Biological significance of RNA editing in cells.

    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

    authors: Tang W,Fei Y,Page M

    更新日期:2012-09-01 00:00:00

  • Responsiveness and Adaptation to Salt Stress of the REDOX-RESPONSIVE TRANSCRIPTION FACTOR 1 (RRTF1) Gene are Controlled by its Promoter.

    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

    authors: Soliman ERS,Meyer P

    更新日期:2019-04-01 00:00:00

  • A highly efficient method for construction of rice artificial MicroRNA vectors.

    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

    authors: Wang X,Yang Y,Yu C,Zhou J,Cheng Y,Yan C,Chen J

    更新日期:2010-11-01 00:00:00

  • Effect of Active Site Pocket Structure Modification of D-Stereospecific Amidohydrolase on the Recognition of Stereospecific and Hydrophobic Substrates.

    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

    authors: Elyas YYA,Miyatani K,Shimizu K,Arima J

    更新日期:2018-09-01 00:00:00

  • Targeted Gene Deletion in Cordyceps militaris Using the Split-Marker Approach.

    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

    authors: Lou H,Ye Z,Yun F,Lin J,Guo L,Chen B,Mu Z

    更新日期:2018-05-01 00:00:00

  • Studies on mRNA electroporation of immature and mature dendritic cells: effects on their immunogenic potential.

    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

    authors: Met O,Eriksen J,Svane IM

    更新日期:2008-10-01 00:00:00

  • A novel strategy for analyzing RNA-protein interactions by surface plasmon resonance biosensor.

    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

    authors: Yang Y,Wang Q,Guo D

    更新日期:2008-09-01 00:00:00

  • Enhancement of Production of D-Glucosamine in Escherichia coli by Blocking Three Pathways Involved in the Consumption of GlcN and GlcNAc.

    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

    authors: Li P,Li X,Wen Z,Dong R,Li K,Fan H,Wang R,Wang J,Lv M

    更新日期:2020-08-01 00:00:00

  • Isolation of the 5'-end of plant genes from genomic DNA by TATA-box-based degenerate primers.

    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

    authors: Guo Y,Ma L,Ji Y,Pu G,Liu B,Du Z,Li G,Ye H,Wang H

    更新日期:2011-02-01 00:00:00

  • Biochemical characterization, molecular cloning, and structural modeling of an interesting β-1,4-glucanase from Sclerotinia sclerotiorum.

    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

    authors: Chahed H,Ezzine A,Mlouka AB,Hardouin J,Jouenne T,Marzouki MN

    更新日期:2014-04-01 00:00:00