A PCR‐RFLP Test to Detect Allelic Variants of the Bovine Kappa‐Casein Gene

Introduction. Numerous molecular genetic studies have revealed a correlation between the polymorphism of milk protein genes and the technological properties of milk raw materials. DNA analysis, in particular, initiated research into the influence of allelic variants of κ-casein (CSN3) on thermal stability and cheese suitability of milk. This gives relevance to our study that compares the results of genotypic identification of lactating cows by the κ-casein gene, using raw and processed milk samples. Study objects and methods. Our study used raw and reconstituted milk samples from first-calf cows of the black motley breed with the AA and BB genotypes of the κ-casein gene. The samples were analyzed by standardized and generally accepted chemical engineering methods, as well as by capillary electrophoresis and PCR-RFLP analysis. Results and discussion. We compared the results of tests on thermal stability and cheese suitability of raw and reconstituted milk samples from cows with the AA and BB genotypes of the κ-casein gene. We tried out an integrated approach to monitoring milk raw materials based on the most relevant technological criteria and correlating the data with the associated CSN3 gene identification parameters. The PCR-RFLP analysis revealed reproducible results for both raw and dried milk samples in relation to the genotypical identification by the A- and B- allelic variants of the CSN3 gene. The tests showed higher thermal stability in the reconstituted milk from the BB genotype cow and better cheese suitability in the AA genotype sample. Conclusion. We developed a system for evaluating milk raw materials based on the most important technological parameters in combination with their genotypic characteristics. Our research procedure can unify the accumulation of experimental data and contribute to the formation of bioinformatics algorithms. This approach can be used in mathematical modeling of criteria to evaluate the compliance of the technological properties of milk with the recommended indicators.

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SLC26A4 Variations Among Graves’ Hyper-Functioning Thyroid Gland

Disease Markers ◽

10.1155/2010/327518 ◽

2010 ◽

Vol 29 (2) ◽

pp. 63-69 ◽

Cited By ~ 7

Author(s):

Hassen Hadj-Kacem ◽

Rihab Kallel ◽

Salima Belguith-Maalej ◽

Mouna Mnif ◽

Ilhem Charfeddine ◽

...

Keyword(s):

Healthy Subjects ◽

Direct Sequencing ◽

Autosomal Recessive Disorder ◽

Hashimoto Thyroiditis ◽

Autoimmune Thyroid Diseases ◽

Allelic Variants ◽

Systematic Analysis ◽

Autoimmune Thyroid ◽

Pcr Rflp ◽

Exon Junctions

Deleterious mutations ofSLC26A4cause Pendred syndrome (PS), an autosomal recessive disorder comprising goitre and deafness with enlarged vestibular aqueducts (EVA), and nonsyndromic hearing loss (NSHL). However, theSLC26A4hyperactivity was recently associated with the emergence of autoimmune thyroid diseases (AITD) and asthma among human and mouse model. Here, by direct sequencing, we investigate the sequences of the 20 coding exons (2 to 21) of SLC26A4 and their flanking intron-exon junctions among patients affected with Graves' disease (GD) hyperthyroidism. Ten mono-allelic variants were identified, seven of which are intronic and previously unreported. Two, c.898A>C (p.I300L) and c.1061T>C (p.F354S), of the three exonic variants are non synonymous. The p.F354S variant is already described to be involved in PS or NSHL inheritances. The exploration by PCR-RFLP of p.I300L and p.F354S variants among 132 GD patients, 105 Hashimoto thyroiditis (HT), 206 Healthy subjects and 102 families with NSHL have shown the presence of both variants. The p.F354S variation was identified both among patients (1~HT and 3 GD) and healthy subjects (n=5). Whereas, the p.I300L variant was identified only in GD patients (n=3). Our studies provide evidence of the importance of systematic analysis ofSLC26A4gene sequences on models other than deafness. This approach allows the identification of new variants and the review of the pathogenic effects of certain mono-allelic variants reported responsible for PS and NSHL development.

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Assessment of Biological Potential Realization in Cattle of the Kostroma Breed with Different Allelic Variants of Beta-Casein Gene

Vestnik APK Verhnevolzh`ia ◽

10.35694/yarcx.2021.54.2.004 ◽

2021 ◽

pp. 22-28

Author(s):

А.А. Чаицкий ◽

Н.С. Баранова

Keyword(s):

Genetic Mutation ◽

Biological Efficiency ◽

Allelic Variants ◽

Casein Gene ◽

Biological Potential ◽

Beta Casein ◽

Casein Protein ◽

High Level ◽

Relationship Of ◽

Quantitative Indicators

Среди сложных белков молока – казеинов – чаще всего встречаются α- и β-казеин. Ген бета-казеина имеет 12 аллельных вариантов, среди которых получили преимущество варианты А1 и А2. При этом, А2 – естественная для человеческого организма разновидность белка бета-казеина, а тип А1 возник в результате природной генетической мутации. Для получения максимального объёма продукции лучшего качества при переориентации производства на молоко А2 необходимо учитывать как основные количественные показатели продуктивности крупного рогатого скота, так и коэффициенты, определяющие эффективность использования коров с точки зрения биологической полноценности продукции, такие как биологической эффективности коров (БЭК), биологической полноценности молока (КБП), кормового коэффициента (FCR) и валовой эффективности использования кормов (GFE). Представлено сравнение количественных и качественных показателей животных разного возраста и групп с учётом степени кровности по улучшающей породе, а также с учётом генотипа по бета-казеину. Впервые проведён анализ реализации биологического потенциала крупного рогатого скота костромской породы с разными аллельными вариантами гена бета-казеина. В ходе исследования установлено, что животные генотипа А2А2 по бета-казеину отличались высоким уровнем БЭК и КБП, а также характеризовались достоверно более высоким уровнем реализации сухого вещества корма в молоко (FCR, GFE), чем у животных А1А2. Так, удои у коров с генотипом А2А2 с кровностью 50% по улучшающей породе за полновозрастную лактацию на 2514 кг больше, чем у животных генотипа А1А2, а коэффициенты БЭК, КБП и FCRвыше на 39,79%, 15,81% и 0,57 кг соответственно. Among the complex proteins of milk – caseins – most often there are α- and β-casein. The beta-casein gene has 12 allelic variants, among which the A1 and A2 variants have gained advantage. At the same time A2 is a natural variety of beta-casein protein for the human body and type A1 arose as a result of a natural genetic mutation. To obtain the maximum volume of production of better quality when reorienting production to milk A2 it is necessary to take into account both the main quantitative indicators of cattle productivity and the coefficients that determine the efficiency of cows use in terms of biological usefulness of products, such as the biological efficiency of cows (BEC), the biological usefulness of milk (BUC), feed coefficient rate (FCR) and gross feed efficiency (GFE). A comparison of quantitative and qualitative indicators of animals of different ages and groups is presented, taking into account the degree of blood relationship by improving breed, as well as taking into account the genotype by beta-casein. For the first time an analysis of the implementation of the biological potential of cattle of the Kostroma breed with different allelic variants of the beta-casein gene was carried out. During the research it was established that animals of a genotype A2A2 on beta casein differed in the BEC and BUC high level and also were characterized by significantly higher level of implementation of dry matter of feed into milk (FCR, GFE), than at animals A1A2. Thus, yields in cows with a genotype A2A2 with a blood relationship of 50% for the improving breed for full-age lactation 2514 kg more than in animals of the genotype A1A2 and the coefficients of BEC, BUC and FCR are higher by 39.79%, 15.81% and 0.57 kg, respectively. Keywords:

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Assessment of polymorphism on kappa-casein gene of Anatolian water buffalo breed using the PCR–RFLP method

TURKISH JOURNAL OF VETERINARY AND ANIMAL SCIENCES ◽

10.3906/vet-2001-3 ◽

2020 ◽

Vol 44 (4) ◽

pp. 904-909

Author(s):

Yusuf ÖZŞENSOY

Keyword(s):

Water Buffalo ◽

Casein Gene ◽

Pcr Rflp

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CCR2, CX3CR1, RANTES and SDF1 genetic polymorphisms influence HIV infection in a Zimbabwean pediatric population

The Journal of Infection in Developing Countries ◽

10.3855/jidc.4599 ◽

2014 ◽

Vol 8 (10) ◽

pp. 1313-1321 ◽

Cited By ~ 3

Author(s):

Kudakwashe Mhandire ◽

Kerina Duri ◽

Gwendoline Kandawasvika ◽

Precious Chandiwana ◽

Nyasha Chin'ombe ◽

...

Keyword(s):

Hiv Infection ◽

Chemokine Receptor ◽

Pediatric Population ◽

Receptor Gene ◽

Allelic Variants ◽

Genotype Combination ◽

High Burden ◽

Pcr Rflp ◽

Minor Alleles ◽

Chemokine Receptor Gene

Introduction: There is growing evidence that polymorphisms in chemokine and chemokine receptor genes influence susceptibility to HIV infection and disease progression. However, not much is documented about the prevalence and effects of chemokine and chemokine receptor gene variations in the Zimbabwean population despite the high burden of HIV/AIDS in the country. This study therefore describes polymorphisms in CCR2, CX3CR1, SDF1 and RANTES genes in a Zimbabwean pediatric population and their effects on HIV infection in children born to HIV-infected mothers. Methodology: A total of 106 children between seven and nine years of age comprising 70 perinatally exposed to HIV (34 born infected [EI] and 36 born uninfected [EU]) and 36 unexposed and uninfected (UEUI) controls were recruited. Six allelic variants in four genes were genotyped using PCR-RFLP and sequencing. Results: Frequencies for minor alleles in the HIV uninfected groups (EU and UEUI) were CCR2 190A (16%), SDF1 801A (2%), CX3CR1 745A (9%), CX3CR1 839T (0%), RANTES In 1.1C (20%), and RANTES -403A (44%). There were significant differences between the EI and EU groups in the distribution of CCR2 190G/A genotype (15% versus 39%, respectively, p = 0.02) and CCR2 190G/A-CX3CR1 745G/G genotype combination (0% versus 33%, respectively, p = 0.002). Conclusions: Our findings suggest that chemokine and chemokine receptor gene variants seem to play an important role in the dynamics of HIV infection and could be used as drug or vaccine targets.

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Polymorphism of the kappa-casein gene in two Bosnian autochthonous cattle breeds

Archives Animal Breeding ◽

10.5194/aab-53-277-2010 ◽

2010 ◽

Vol 53 (3) ◽

pp. 277-282

Author(s):

M. Brka ◽

A. Hodžić ◽

N. Reinsch ◽

E. P. Zečević ◽

A. Dokso ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Restriction Fragment Length Polymorphism ◽

Fragment Length Polymorphism ◽

Small Population ◽

Chain Reaction ◽

Casein Gene ◽

Pcr Rflp ◽

Polymerase Chain ◽

Related Breed ◽

Restriction Fragment Length

Abstract. Buša is an old endangered autochthonous breed of the western Balkan, especially Bosnia-Herzegovina, Kosovo and Albania. A related breed is Gatačko, derived from Buša × Tirolean Grey crossbreds. Fifteen purebred Buša cattle and thirteen Gatačko animals were genotyped for polymorphisms at the kappa-casein gene by a Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) essay. The alleles A, B and C were found and the allelic frequencies were 0.46 (A), 0.46 (B) and 0.08 (C) in Buša cattle and 0.58 (A) and 0.42 (B) in Gatačko. Only AA, AB, BB and BC genotypes occurred. Further alleles were not detected and are therefore either absent in both populations or rare. The allele »B« found in this small population will be useful for a sire selection program in the future.

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Genetic polymorphism of kappa-casein gene in Friesian Holstein: a basic selection of dairy cattle superiority

Journal of the Indonesian Tropical Animal Agriculture ◽

10.14710/jitaa.42.4.213-219 ◽

2017 ◽

Vol 42 (4) ◽

pp. 213 ◽

Cited By ~ 4

Author(s):

S. D. Volkandari ◽

I. Indriawati ◽

E. T. Margawati

Keyword(s):

Dairy Cattle ◽

Milk Protein ◽

Casein Gene ◽

Genotype Frequencies ◽

Pcr Rflp ◽

Milk Component ◽

Milk Performance ◽

Pcr Method ◽

Two Populations ◽

Selection Of

Caseins are milk protein subdivided into four main groups which are αS1, αS2, β-casein and kappa-casein (CSN3). Kappa-casein gene influences the manufacturing of milk properties. The aim of this study was to identify the kappa-casein gene polymorphism in Friesian Holstein (FH) cattle. Fifty nine (59) samples consisted of 32 (Malang), 10 (Sukahati Bogor) and 17 (Research Center for Biotechnology-Indonesian Institute of Sciences’s collections)were applied in this study. DNA samples were extracted by high concentrated NaCl and quantified by spectrophotometer. The kappa-casein gene was amplified at 379 bp fragment by PCR method using a pair primer of kappa-casein at 56oC annealing for 30 cycles. PCR-RFLP technique with HindIII was used for genotyping analysis. The result showed that there were three variants of genotypes (AA, AB and BB) in two populations from Malang and RC for Biotechnology-LIPI’s collection while cattle from Sukahati had only AA and AB genotypes. The averages of genotype frequencies were 65.28%; 65.28%; and 3.00% for AA, AB and BB genotypes respectively while frequencies of 0.81 and 0.19 were for A and B alleles, respectively. FH cattle populations were in equilibrium genetics. This finding concludes that polymorphism was found in three of FH populations with A allele was more common in kappa-casein locus. B allele is known having association with milk production, milk component and cheese yield. Increasing of B allele would influence on milk performance of FH cattle. Explorations of quantitative, qualitative and molecular genetics are important to improve dairy cattle performance.

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Inheritance of allelic variants of the kappa-casein gene by cows

BIO Web of Conferences ◽

10.1051/bioconf/20201700059 ◽

2020 ◽

Vol 17 ◽

pp. 00059 ◽

Cited By ~ 3

Author(s):

Radik Shaidullin ◽

Gazimzyan Sharafutdinov ◽

Anastasia Moskvicheva ◽

Tagir Faizov ◽

Yusupzhan Yuldashbaev

Keyword(s):

Milk Yield ◽

Milk Fat ◽

Mass Fraction ◽

Milk Protein ◽

Frequency Of Occurrence ◽

Allelic Variants ◽

Casein Gene ◽

Black And White ◽

White Cattle ◽

Pcr Diagnostics

The inheritance of allelic variants of the kappa-casein gene from bulls with different genotypes and their influence on the milk production of daughters were studied. For research, PCR diagnostics of daughters in the herds of black-and-white cattle at the farms “Azeleevo” and “Vakhitovo” (OJSC “Red East Agro”) was carried out, the fathers were two bulls LADDIE 135797213 with a genotypes CSN3 AB and LOMAX 10785322 with a genotype CSN3 BB. Based on the results of genotyping, the frequency of occurrence of genotypes AA, AB, BB kappa-casein and allelic variants A and B in daughters was calculated. The bull with the CSN3 AB genotype produced the largest number of offspring with the CSN3 AA genotype 50.0-51.8 %, and the bull with the homozygous BB genotype produced 24–25 % of daughters carrying the BB variant and 75 -76 % of the AB genotype. The frequency of occurrence of the Kappa-casein allele in the daughters of bulls with the genotype CSN3 BB was 0.63, which is almost 2 times more than the allele A. When comparing heifers with the same genotype (CSN3 AB), it was found that at the Azeleevo farm, the daughters of the bulls having the genotype BB of kappa-casein are superior to cows born from bulls with the genotype AB by the milk yield (+36 kg), mass fraction of fat and protein (+0.01 %), the amount of milk fat (+2 kg) and protein (+2 kg). In the group of first-calf heifers with the BB genotype, daughters of the producer with the BB genotype are superior by the milk yield (+ 175 kg), the mass fraction of fat in milk (+0.1 %), the amount of milk fat (+12 kg), the mass fraction of protein in milk (+0.09 %), the amount of milk protein (+11 kg). At the Vakhitovo farm, similar results in the group of CSN3 AB heifers born from the bulls with the BB genotype was observed

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New determination method for the ratio of the relative proportions of ϰ-casein alleles in milk powder

Food Processing Techniques and Technology ◽

10.21603/2074-9414-2020-3-525-535 ◽

2020 ◽

Vol 50 (3) ◽

pp. 525-535

Author(s):

Khamid Gilmanov ◽

Vladislav Semipyatniy ◽

Alana Bigaeva ◽

Ramil Vafin ◽

Svetlana Turovskaya

Keyword(s):

Dairy Products ◽

Raw Materials ◽

Milk Powder ◽

Rflp Analysis ◽

Raw Milk ◽

Raw Material ◽

Technological Properties ◽

Additional Indication ◽

Casein Gene ◽

Pcr Rflp

Introduction. High-quality dairy products depend on the properties of raw materials. Milk is the basic raw material for all dairy products. Its quality is subject to requirements of biological safety, and its physicochemical and technological properties comply with strict indicators. ϰ-casein gene is a basic milk protein. Its allelic variants affect milk productivity, as well as its composition and technological properties, e.g. cheeseability and heat resistance. The research objective was to develop a new method for determining the ratio of the relative proportions of selection-significant kappa-casein gene alleles in bulk cow’s milk powder by means of molecular genetics and bioinformation system. The method can help to test raw materials for further processing rationality. Study objects and methods. The research included a genetic analysis of samples of bulk whole milk powder, bulk powdered skimmed milk, and bulk raw milk. The developed method involved DNA extraction, combined PCR-RFLP technique, electrophoretic results, and analysis detection of information data using new mathematical algorithms and software. Results and discussion. The ratio of the relative proportions of the kappa-casein gene alleles in milk powder was determined using primer sets JK5 and JK3, as well as restriction endonuclease HinfI for PCR-RFLP analysis. The experiment showed satisfactory reproducibility and interpretation of the obtained data. The program provided a ratio calculation of the kappa-casein gene alleles relative proportions in the studied milk powder samples, expressed as a percentage of the A allele proportion with additional indication of the absolute and relative errors. It also placed the information block of the generated numerical indicators into the percentage scale. Conclusion. The developed methodology was implemented by a set of laboratory procedures. The new specialized program “Calculation of the relative proportions ratio of ϰ-casein alleles in bulk milk” is on open access. It provides a correct and prompt data interpretation, generated during the analysis of dry milk raw materials of bulk origin.

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