scholarly journals DNA authentication of brewery products: basic principles and methodological approaches

2019 ◽  
pp. 364-374 ◽  
Author(s):  
Lev Oganesyants ◽  
Ramil Vafin ◽  
Aram Galstyan ◽  
Anastasia Ryabova ◽  
Sergey Khurshudyan ◽  
...  
Keyword(s):  
Hordeum Vulgare ◽  
Nucleic Acids ◽  
Dna Extraction ◽  
Nuclear Dna ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Snp Markers ◽  
Pcr Analysis ◽  
Malting Barley ◽  
Barley Malt

Beer DNA authentication is the process of authentication by identification of barley malt Hordeum vulgare or its substitutes, as well as hops and yeast. The method is based on molecular genetic analysis of residual quantities of nucleic acids extracted from the cellular debris of the final product. The aim of the study was to analyse scientific and methodical approaches to extraction of residual quantities of beer raw materials nucleic acids and beer DNA authentication for their later application in determining brewing products authenticity. The technological level discloses the method of DNA extraction from wines, modified for extraction of nucleic acids from beer samples. The method includes the following characteristic peculiarities: stage enzymatic hydrolysis of polysaccharides and polypeptides of dissolved lyophilisate, multiple sedimentation and resursuspension of nucleoproteid complex, RNA removal followed by DNA extraction by organic solvents, and additional DNA purification by magnetic particle adsorption. This review presents the analysis of genetic targets used as molecular markers for gene identification of malting barley varieties and beer DNA authentication. We also provided the interpretation of PCR analysis of Hordeum vulgare varieties and samples of commercial beer. Data on SSR- and SNP-markers of Hordeum vulgare nuclear DNA, used for barley varieties identification and potentially suitable for beer DNA authentication, are also presented. We also analysed genetic targets used in malting barley substitute detection, as well as hops and yeast identification in beer. Data on correlation of amplified DNA targets with beer quality indicators were systematised.

scholarly journals Introduction to Molecular Diagnostics of Insects

2021 ◽  
Vol 104 (4) ◽  
pp. 184-195
Author(s):  
А. S. Ryabinin* ◽  
R. А. Bykov ◽  
V. К. Lapshina ◽  
А. А. Maslakova ◽  
М. А. Demenkova ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Traditional Approach ◽  
Molecular Genetic ◽  
General Information ◽  
Molecular Genetic Analysis ◽  
Molecular Methods ◽  
Pcr Analysis ◽  
Morphological Studies ◽  
Dna Quality ◽  
Wide Range

Insects play an important role in biocenoses due to their abundance and wide (cosmopolitan) distribution. Many insects are crop pests. An effective pest control could be realized in case of proper species identification, which is usually managed by morphological analysis. Molecular methods allow to deep study of many issues of insect biology. In particular, traditional approach can not ordinary identify a species at all stages of their life cycle, whereas molecular methods can it. This review covers a wide range of issues related to the molecular genetic analysis of insects. In the first section we consider the methods of fixation and storage of insect specimens, as well as their impact on DNA quality. Further, we provide general information on population study design. Various schemes of DNA extraction, examples of both express techniques and more thorough protocols for DNA extraction and their purification are provided. In addition, methods of DNA isolation that allow to preserve a specimen integrity for further morphological studies are considered. The methods of DNA quality control are described in detail, that is important for PCR analysis. The last section provides various methods of PCR analysis, that we exemplify by studies aimed to elucidate both fundamental issues and practical problems.

Molecular Genetic Analysis of a Compound Heterozygote for the Glycoprotein (GP) IIb Gene Associated With Glanzmann’s Thrombasthenia: Disruption of the 674-687 Disulfide Bridge in GPIIb Prevents Surface Exposure of GPIIb-IIIa Complexes

Blood ◽  
1999 ◽  
Vol 93 (3) ◽  
pp. 866-875 ◽  
Author(s):  
Consuelo González-Manchón ◽  
Marta Fernández-Pinel ◽  
Elena G. Arias-Salgado ◽  
Milagros Ferrer ◽  
M.-Victoria Alvarez ◽  
...  
Keyword(s):  
Molecular Genetic ◽  
Splice Junction ◽  
Disulfide Bridge ◽  
Molecular Genetic Analysis ◽  
Pcr Analysis ◽  
Polymorphism Analysis ◽  
Glanzmann’S Thrombasthenia ◽  
Single Stranded Conformational Polymorphism ◽  
Glanzmann's Thrombasthenia ◽  
Platelet Content

Abstract This work was aimed at elucidating the molecular genetic lesion(s) responsible for the thrombasthenic phenotype of a patient whose low platelet content of glycoprotein (GP) IIb-IIIa indicated that it was a case of type II Glanzmann’s thrombasthenia (GT). The parents did not admit consanguinity and showed a reduced platelet content of GPIIb-IIIa. Polymerase chain reaction (PCR)–single-stranded conformational polymorphism analysis of genomic DNA showed no mutations in the patient’s GPIIIa and two novel mutations in the GPIIb gene: one of them was a heterozygous splice junction mutation, a C→A transversion, at position +2 of the exon 5-intron 5 boundary [IVS5(+2)C→A] inherited from the father. The predicted effect of this mutation, insertion of intron 5 (76 bp) into the GPIIb-mRNA, was confirmed by reverse transcription-PCR analysis of platelet mRNA. The almost complete absence of this mutated form of GPIIb-mRNA suggests that it is very unstable. Virtually all of the proband’s GPIIb-mRNA was accounted for by the allele inherited from the mother showing a T2113→C transition that changes Cys674→Arg674 disrupting the 674-687 intramolecular disulfide bridge. The proband showed a platelet accumulation of proGPIIb and minute amounts of GPIIb and GPIIIa. Moreover, transfection and immunoprecipitation analysis demonstrated that [Arg674]GPIIb is capable of forming a heterodimer complex with GPIIIa, but the rate of subunit maturation and the surface exposure of GPIIb-IIIa are strongly reduced. Thus, the intramolecular 674-687 disulfide bridge in GPIIb is essential for the normal processing of GPIIb-IIIa complexes. The additive effect of these two GPIIb mutations provides the molecular basis for the thrombasthenic phenotype of the proband.

Molecular Genetic Analysis of a Compound Heterozygote for the Glycoprotein (GP) IIb Gene Associated With Glanzmann’s Thrombasthenia: Disruption of the 674-687 Disulfide Bridge in GPIIb Prevents Surface Exposure of GPIIb-IIIa Complexes

Blood ◽  
1999 ◽  
Vol 93 (3) ◽  
pp. 866-875 ◽  
Author(s):  
Consuelo González-Manchón ◽  
Marta Fernández-Pinel ◽  
Elena G. Arias-Salgado ◽  
Milagros Ferrer ◽  
M.-Victoria Alvarez ◽  
...  
Keyword(s):  
Molecular Genetic ◽  
Splice Junction ◽  
Disulfide Bridge ◽  
Molecular Genetic Analysis ◽  
Pcr Analysis ◽  
Polymorphism Analysis ◽  
Glanzmann’S Thrombasthenia ◽  
Single Stranded Conformational Polymorphism ◽  
Glanzmann's Thrombasthenia ◽  
Platelet Content

This work was aimed at elucidating the molecular genetic lesion(s) responsible for the thrombasthenic phenotype of a patient whose low platelet content of glycoprotein (GP) IIb-IIIa indicated that it was a case of type II Glanzmann’s thrombasthenia (GT). The parents did not admit consanguinity and showed a reduced platelet content of GPIIb-IIIa. Polymerase chain reaction (PCR)–single-stranded conformational polymorphism analysis of genomic DNA showed no mutations in the patient’s GPIIIa and two novel mutations in the GPIIb gene: one of them was a heterozygous splice junction mutation, a C→A transversion, at position +2 of the exon 5-intron 5 boundary [IVS5(+2)C→A] inherited from the father. The predicted effect of this mutation, insertion of intron 5 (76 bp) into the GPIIb-mRNA, was confirmed by reverse transcription-PCR analysis of platelet mRNA. The almost complete absence of this mutated form of GPIIb-mRNA suggests that it is very unstable. Virtually all of the proband’s GPIIb-mRNA was accounted for by the allele inherited from the mother showing a T2113→C transition that changes Cys674→Arg674 disrupting the 674-687 intramolecular disulfide bridge. The proband showed a platelet accumulation of proGPIIb and minute amounts of GPIIb and GPIIIa. Moreover, transfection and immunoprecipitation analysis demonstrated that [Arg674]GPIIb is capable of forming a heterodimer complex with GPIIIa, but the rate of subunit maturation and the surface exposure of GPIIb-IIIa are strongly reduced. Thus, the intramolecular 674-687 disulfide bridge in GPIIb is essential for the normal processing of GPIIb-IIIa complexes. The additive effect of these two GPIIb mutations provides the molecular basis for the thrombasthenic phenotype of the proband.

scholarly journals DNA from mollusc shell: a valuable and underutilised substrate for genetic analyses

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9420 ◽  
Author(s):  
Sara Ferreira ◽  
Rachael Ashby ◽  
Gert-Jan Jeunen ◽  
Kim Rutherford ◽  
Catherine Collins ◽  
...  
Keyword(s):  
Success Rate ◽  
Nuclear Dna ◽  
Molecular Genetic ◽  
Nuclear Gene ◽  
Molecular Genetic Analysis ◽  
Ecological Communities ◽  
Mollusc Shell ◽  
Genetic Analyses ◽  
Perna Canaliculus ◽  
Beach Cast

Mollusc shells are an abundant resource that have been long used to predict the structures of ancient ecological communities, examine evolutionary processes, reconstruct paleoenvironmental conditions, track and predict responses to climatic change, and explore the movement of hominids across the globe. Despite the ubiquity of mollusc shell in many environments, it remains relatively unexplored as a substrate for molecular genetic analysis. Here we undertook a series of experiments using the New Zealand endemic greenshell mussel, Perna canaliculus, to explore the utility of fresh, aged, beach-cast and cooked mollusc shell for molecular genetic analyses. We find that reasonable quantities of DNA (0.002–21.48 ng/mg shell) can be derived from aged, beach-cast and cooked mussel shell and that this can routinely provide enough material to undertake PCR analyses of mitochondrial and nuclear gene fragments. Mitochondrial PCR amplification had an average success rate of 96.5% from shell tissue extracted thirteen months after the animal’s death. A success rate of 93.75% was obtained for cooked shells. Amplification of nuclear DNA (chitin synthase gene) was less successful (80% success from fresh shells, decreasing to 10% with time, and 75% from cooked shells). Our results demonstrate the promise of mollusc shell as a substrate for genetic analyses targeting both mitochondrial and nuclear genes.

scholarly journals Genetic and physiological analysis T1 biotechnological plants of winter wheat (Triticum aestivum L.)

2020 ◽  
Vol 26 ◽  
pp. 222-227
Author(s):  
A. G. Komisarenko ◽  
S. I. Mykhalska ◽  
V. M. Kurchii ◽  
O. O. Khrystan
Keyword(s):  
Triticum Aestivum ◽  
Winter Wheat ◽  
First Generation ◽  
Genetically Modified ◽  
Molecular Genetic ◽  
Triticum Aestivum L ◽  
Molecular Genetic Analysis ◽  
Pcr Analysis ◽  
Proline Dehydrogenase ◽  
In Planta

Aim. To investigate inheritance of transgenes in the first generation (T1) of winter wheat biotechnological plants (Triticum aestivum L.). Analyze the performance of T1 genetically modified plants with a double stranded RNA suppressor of the proline dehydrogenase (pdh) gene under normal growing conditions. Methods. PCR analysis, DNA electrophoresis; determination of indicators of the structure of the crop. Results. Molecular genetic analysis was performed and the performance indicators of control and T1 biotechnological plants were investigated. Conclusions. The first generation of genetically modified winter wheat plants resulting from Agrobacterium-mediated transformation in planta confirmed the inheritance of integrated genes. Among the transgenic variants identified plants that lack some fragments of the target gene required for partial suppression of the gene of proline dehydrogenase wheat. It is shown that at the optimal terms of growing biotechnological plants of wheat winter-annual of UK 322/17 and UK 209 h was characterized by the best indexes of structure of harvest as compared to an initial form, while the genetically changed plants of genotypes of UK 95/17 and UK 065 after the elements of the productivity did not differ from control plants. Keywords: Triticum aestivum L., biotechnological plants, T-DNA, proline dehydrogenase gene, structural analysis indicators.

scholarly journals Prospects for DNA authentication in wine production monitoring

2018 ◽  
Vol 6 (2) ◽  
pp. 438-448 ◽  
Author(s):  
Lev Oganesyants ◽  
Lev Oganesyants ◽  
Ramil Vafin ◽  
Ramil Vafin ◽  
Aram Galstyan ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Vitis Vinifera ◽  
Raw Materials ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Genetic Identification ◽  
Vitis Vinifera L ◽  
Wine Production ◽  
Dna Yield

Wines DNA authentication is a technological process of their authenticity verification by genetic identification of the main plant ingredient by means of molecular genetic analysis of the residual amounts of Vitis vinifera L nucleic acids extracted from end product cellular debris. The main aim of the research was the analysis of scientific and methodological approaches to the extraction of residual amounts of nucleic acids in wine raw materials and DNA authentication of wines for their subsequent application in solving the problem of determining wine products authenticity and place of origin. The prior art includes various approaches to the extraction of Vitis vinifera L. nucleic acids among which the three methods by Savazzini & Martinelli, Pereira and Bigliazzi can be named basically. Analysis of the effectiveness of different methods of DNA extraction from wines indicates the superiority of the Pereira method over other traditional methods of extraction in terms of DNA yield and quality. Besides, the nucleic acid extracted from wines is characterized as residual since its concentration is significantly reduced in a multi-stage wine production process. The yield of extracted nucleic acid also decreases as the wine ages. The use of microsatellite DNA loci designed for grapes genetic identification is one of the approaches applicable for wine DNA authentication.

scholarly journals INFLUENCE OF BLOOD GROUPS, CAST AND BLG GENES ON PRODUCTIVITY OF THE SHEEP AND GOATS OF THE ALTAY REPUBLIC

2018 ◽  
Vol 48 (4) ◽  
pp. 63-71
Author(s):  
G. M. Goncharenko ◽  
N. V. Grishina ◽  
T. S. Khoroshilova ◽  
I. V. Romanchuk ◽  
T. B. Kargachakova ◽  
...  
Keyword(s):  
Genetic Similarity ◽  
Molecular Genetic ◽  
Live Weight ◽  
Molecular Genetic Analysis ◽  
Pcr Analysis ◽  
Dna Diagnosis ◽  
Genetic Characteristics ◽  
High Productivity ◽  
The Republic ◽  
Cast Gene

The use of genetic markers in addition to traditional methods of animal selection with desirable genotypes allows to increase the share of animals with high productivity in the next generations and ensures improvement of breeding efficiency. Genetic features of the Prikatun type of the Gorno-Altay semi-fine wool breed of sheep and the white downy goat breed in the Republic of Altay were studied by the method of immunogenetic and molecular genetic analysis. The frequency of antigenic factors was identified and the index of genetic similarity between goats and sheep and their separate herds was calculated. Gene polymorphism of β-lactoglobulin (BLG) and calpastatin (CAST) was revealed by the method of DNA diagnosis. Population and genetic characteristics of the herds was studied by the genes specified. Associative genotype relation to productivity and quality of the produce obtained was analyzed. The index of genetic similarity between the goats and the sheep was at the level of 0.713, between the separate herds of the goats the index was 0.861. The ratio of genotypes in the BLG gene determined by PCR analysis in the white downy goats was S1S1– 16.1%; S1S2– 50.6%; S2S2– 33.3%. In the Prikatun type two genotypes were identified in this gene: BB with the frequency of 59.2%, and AB – 40.8%. Two different alleles were identified in the CAST gene of sheep (M and N). The genotype MM was the predominant variant in the CAST sheep gene, whose frequency was 88%. The frequency of occurrence of animals with NN genotype was 1%, MN – 11%. It was shown that the gene equilibrium in the herds was not broken, χ2= 0.931. It was noted that heterozygous goats (S1S2) by BLG gene had a higher live weight by 0.30-0.61 kg compared to other variants of BLG gene (p<0.05). It was also found that lambs with genotype MM of the CAST gene had a higher live weight by 5.5 kg than MN heterozygotes (p< 0.01). However, this difference was not revealed in other age and sex groups of animals.

scholarly journals Biological Properties and Genetic Characteristics of Francisella tularensis Strains Isolated in the Territory of the Rostov Region in 2020

2021 ◽  
pp. 134-140
Author(s):  
M. V. Tsimbalistova ◽  
V. M. Sorokin ◽  
N. V. Aronova ◽  
A. S. Anisimova ◽  
N. L. Pichurina ◽  
...  
Keyword(s):  
Francisella Tularensis ◽  
Molecular Genetic ◽  
Biological Properties ◽  
Snp Markers ◽  
Pcr Analysis ◽  
Snp Analysis ◽  
Genetic Characteristics ◽  
Tularensis Subsp ◽  
Rostov Region ◽  
Natural Foci

Objective of the study was to investigate biological properties and genetic characteristics of tularemia agent strains isolated from natural foci of the Rostov Region in 2020.Materials and methods. Field material from natural foci of the Rostov Region was examined by serological, bacteriological, biological, and molecular-genetic methods. Cultural-morphological, biochemical, antigenic and pathogenic properties of isolated cultures were studied. Protein profles were obtained through MALDI-TOF MS using mass spectrometer Autoflex speed III Bruker Daltonics and Flex Control of Biotyper software. The genetic characteristics of the strains were determined by VNTR and INDEL typing and SNP analysis.Results and discussion. Six strains of tularemia pathogen were isolated from mouse-like rodents using biological method. The investigation of their biological features and data of PCR analysis and INDEL typing with canonical markers showed that all strains are typical representatives of the Francisella tularensis subsp. holarctica biovar EryR. VNTR typing by six genetic loci revealed that all strains belong to four individual genotypes. The strain isolated in 2020 in the Salsky district was identical to the strain which was isolated in the same area in 1989. Based on the whole genome sequencing of two strains, we established that they are closest to the cultures isolated in Turkey (2009, 2012) and Khanty-Mansiysk (2013) by the studied set of SNP markers. Thus, we found that both identical (or closely related) clones of the tularemia agent and new strains with unique genotypes which previously were not described for the Rostov Region can circulate in natural foci of this region for a long period of time.

scholarly journals KASPar SNP genetic map of cassava for QTL discovery of productivity traits in moderate drought stress environment in Africa

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Favour Ewa ◽  
Joseph N. A. Asiwe ◽  
Emmanuel Okogbenin ◽  
Alex C. Ogbonna ◽  
Chiedozie Egesi
Keyword(s):  
Crop Improvement ◽  
Molecular Genetic ◽  
Complex Trait ◽  
Interval Mapping ◽  
Growth Cycle ◽  
Molecular Genetic Analysis ◽  
Snp Markers ◽  
Genetic Dissection ◽  
Molecular Tools ◽  
High Performing

AbstractCassava is an important staple in Sub-Sahara Africa. While its production has rapidly expanded to the dry savannahs of the continent, productivity is low in this ecology due to drought by farmers, extending the growth cycle from 12 months to 18, and sometimes 24 months to ensure better harvests. Yield is a complex trait and often difficult to manipulate for genetic gain in conventional breeding. Unfortunately, the dearth of molecular tools for decades has hampered molecular breeding (MB) to improve cassava productivity. This study was conducted to explore KASpar SNPs to generate more molecular tools to enhance genetic dissection of elite African germplasm for improved cassava productivity in dry environments of Africa where molecular resources are highly limited for crop improvement. To aid molecular genetic analysis of traits, a linkage map covering 1582.8 cM with an average resolution of 3.69 cM was constructed using 505 polymorphic SNP markers distributed over 21 linkage groups. Composite interval mapping using 267 F1 progeny in initial QTL mapping identified 27 QTLs for productivity traits in the dry savannah of Nigeria. The availability of KASPar SNPs are anticipated to improve the implementation of MB for the development of high performing drought-tolerant cassava varieties in Africa.

A Method for Processing Digital Images of Colorimetric Biochipes for Quantitative Determination for Bacterial Antibiotic Resistance

2021 ◽  
Vol 37 (5) ◽  
pp. 123-131
Author(s):  
G.V. Presnova ◽  
V.G. Grigorenko ◽  
M.M. Ulyashova ◽  
М.Yu. Rubtsova
Keyword(s):  
Antibiotic Resistance ◽  
Nucleic Acids ◽  
Quantitative Determination ◽  
Resistance Genes ◽  
Molecular Genetic ◽  
Antibiotic Resistance Genes ◽  
Molecular Genetic Analysis ◽  
Beta Lactamases ◽  
The Government

Abstract-Molecular genetic analysis methods based on the technology of colorimetric biochip have shown their effectiveness in identifying antibiotic resistance genes in bacteria. For the quantitative determination of nucleic acids, a comparative study of methods for converting digital color images of biochips into monochrome black-and-white versions using RGB and CMYK color models has been carried out. A 19-mer single-stranded oligonucleotide and two model mRNAs corresponding to the genes of two types of clinically relevant beta-lactamases (CTX-M and NDM) were studied as objects. The widest range of staining intensity and the best analytical characteristics for the determination of all types of studied nucleic acids were obtained using the red channel of the RGB color model. The detection limits were 0.10 ± 0.02 pmol/μl for the 19-mer oligonucleotide, and 3.0 ± 0.2 amol/μl and 8.0 ± 0.6 amol/μl for mRNA of beta-lactamases CTX-M-116 and NDM-1, respectively. The developed method can be used for the quantitative determination of expressing antibiotic resistance genes in bacteria with multiple resistance to antimicrobial drugs. Key words: colorimetric biochips, hybridization analysis, DNA, mRNA, antibiotic resistance, beta-lactamases The work was supported by the Government Program of the Lomonosov Moscow State University (АААА-А21-121011290089-4).

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