A method for the simulation of normal, carrier and affected controls for PCR-RFLP screening of a genetic disease in dairy cattle

Abstract. Research to assess the effect of single genes on reproductive traits in bovine species is imperative to elucidate genes' functions and acquire a better perspective of quantitative traits. The present study was undertaken to characterize genetic diversity in the bovine growth hormone (GH) gene in a population of 410 Holstein dairy cows in Tunisia. The analyses were based on single nucleotide polymorphisms, and GH-AluI and GH-MspI detections and genotyping were carried out using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Data were analyzed using a mixed linear model with the MIXED procedure to reveal the possible effect of GH genotypes on reproductive traits. The frequency data of AluI(L//V) and MspI(+//−) alleles were 87.04//12.96 and 70.06//29.94, respectively. The distribution of the frequency of GH genotypes for LL/LV/VV and (−/−)//(+/−)//(+/+) were 77.75//18.59//3.66 and 15.37//29.13//55.50, respectively. The results of the statistical analyses proved that GH-AluI showed a substantial favorable effect on exanimate traits except for the age at first calving; however, only a suggestive effect of GH-MspI on the calving interval (CI) and the days open (DI) was found. The homozygous LL genotype seemed to be advantageous with respect to the CI and the DI compared with LV and VV genotypes. Heterozygous MspI(+/−) cows tended to have a longer CI and DI than MspI(+/+) and MspI(−/−) cows, but the difference was not statistically significant. A significant effect of different GH-AluI–MspI combined genotypes was found on the number of inseminations per conception, the CI and the DI, and the LL/– combined genotype seemed to be associated with better reproductive performance. Based on these results, the LL genotype of the GH locus can be considered to be a favorable genotype for reproductive traits in Holstein dairy cattle, although these findings need to be confirmed by further research before polymorphisms can be used in a marker-assisted selection program.

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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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Effect of beta lactoglobulin polymorphism on milk production in Holstein dairy cattle of Iran

Proceedings of the British Society of Animal Science ◽

10.1017/s1752756200019104 ◽

2007 ◽

Vol 2007 ◽

pp. 7-7

Author(s):

Sonia Zakizadeh ◽

Monika Reissmann ◽

Peter Reinecke ◽

Ghodrat Rahimi ◽

Ardeshir Nejati-Javaremi ◽

...

Keyword(s):

Dairy Cattle ◽

Milk Production ◽

Allelic Frequency ◽

Major Protein ◽

Fat Percentage ◽

Common Genetic Variants ◽

Pcr Rflp ◽

Total Milk ◽

Beta Lactoglobulin ◽

Positive Effect

Beta-Lacto globulin (BLG) is a major protein in the milk of ruminants. Bovine BLG gene is located on the chromosome 11q28 and has two common genetic variants, ‘A’ and ‘B’ (Gene-Bank X14710). These variants are different in amino acids at positions 64 and 118, where allele ‘A’ has Asp and Val and allele ‘B’ has Gly and Ala, respectively. Difference at position 118 can be detected by PCR-RFLP (Medrano and Agiular-Cordova 1990). It is reported that allele ‘A’ is associated with milk production (Strzalkowska et al. 2002) and allele ‘B’ has a positive effect on fat percentage (Tsiaras et al. 2005). The aim of our study was to estimate the allelic frequency in polymorphic site of exon IV (allele ‘A’ or HaeIII (-)) of BLG gene in Holstein dairy cattle of Iran and also to find any association of total milk production (first lactation) with BLG genotypes.

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Genotyping of the polymorphism within exon 3 of prolactin gene in various dairy breeds by PCR RFLP (Brief report)

Archives Animal Breeding ◽

10.5194/aab-51-298-2008 ◽

2008 ◽

Vol 51 (3) ◽

pp. 298-299 ◽

Cited By ~ 2

Author(s):

A. Ratna Kumari ◽

K. M. Singh ◽

K. J. Soni ◽

R. K. Patel ◽

J. B. Chauhan ◽

...

Keyword(s):

Mammary Gland ◽

Dairy Cattle ◽

Genetic Characterization ◽

Bos Taurus ◽

Bos Indicus ◽

Genotype Frequencies ◽

Prolactin Gene ◽

Pcr Rflp ◽

Transition Mutation

Abstract. In mammals, especially dairy cattle the prolactin has important functions like the development of mammary gland affecting milk yield and composition. It has been mapped to chromosome 23 in Bovine (HALLERMAN et al., 1988). A silent A→G transition mutation at the codon for amino acid 103 in exon 3 of bovine prolactin (bPRL) gene gives rise to a polymorphic Rsa I site, has become a popular genetic marker used for genetic characterization of cattle populations by means of PCR-RFLP (MITRA et al., 1995; CHRENEK et al., 1998; DYBUS, 2005). The present study reports on the genotype frequencies observed in various Bos taurus and Bos indicus dairy cattle breeds.

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Characteristics of the genetic structure of bulls of lebedinian breed by Beta (CSN2) - and Capa Casein genes (CSN3)

145 - Tehnologìâ virobnictva ì pererobki produktìv tvarinnictva ◽

10.33245/2310-9289-2020-158-2-88-96 ◽

2020 ◽

pp. 88-96

Author(s):

V. Ladyka ◽

Y. Sklyarenko ◽

Y. Pavlenko

Keyword(s):

Dairy Cattle ◽

Animal Husbandry ◽

Raw Milk ◽

Adaptive Capabilities ◽

Pcr Rflp ◽

Beta Casein ◽

Hereditary Factors ◽

Genetic Level ◽

Casein Genes ◽

Selection Of

The Conservation of genetic resources in animal husbandry is a necessity arising from the possibilities and reality of the diversity of the gene pool available in the country, which determines the productive and adaptive capabilities of individual breeds of cattle. Aboriginal breeds are carriers of unique genes and gene complexes, which cannot be restored if they disappear. Marker-asissted selection is one of the current areas of improvement of dairy cattle productivity. It allows the selection of parental forms at the genetic level. An important issue in the selection of dairy cattle today is the study of the relationship between hereditary factors that determine the types of proteins in milk. Genetic variants of beta-casein significantly affect on human health, kappa-casein is associated with the quality of raw milk and cheese making properties. The aim of the research was to study the available sperm production of Lebedinian bulls by beta- (CSN2) and kappacasein (CSN3) genes. The material for research was the sperm of bulls of Lebedinian breed cattle. Determination of beta- and kappa-casein gene polymorphism was performed by PCR-RFLP method in the laboratory of the Institute of Animal Husbandry of NAAS. Analysis of the genealogical structure showed that 12 bulls-producers belong to 8 lines. Of the 12 bulls, 5 are purebred Lebedinian, 7 are crossbreeds with the swiss breed. Among those studied for the complex genotype (CSN2- (CSN3), one bull had the desired genotype A2A2BB, another bulls had genotypes A2A2AB; A1A2BB; A1A2AB; A1A1AA. Five bulls have the A1A2AA genotype and two bulls have the A1A1AB genotype. The high frequency of the A2 allele of beta-casein (46%) and the B allele of kappa-casein (33%) was observed in sires of Lebedinian breed. To create micropopulations with the desired complex A2A2 / BB genotype, sires are recommended in which the frequency of alleles of beta-casein A2 and kappacasein B satisfies the requirements. Key words: sire, beta-casein, kappa-casein, genotype, allele, frequency, haploid.

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Study association of bPRL (Bovine Prolactin) gene and milk production trait in Indonesian Holstein dairy cattle

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/902/1/012007 ◽

2021 ◽

Vol 902 (1) ◽

pp. 012007

Author(s):

B Hifni ◽

G Pambuko ◽

R E P Lestari ◽

N Herowati ◽

R Vanessa ◽

...

Keyword(s):

Dairy Cattle ◽

Milk Production ◽

Milk Production Trait ◽

Production Traits ◽

Product Size ◽

Pcr Product ◽

Prolactin Gene ◽

Pcr Rflp ◽

Genotype Difference ◽

Study Association

Abstract This study was aimed to analyze the genetic variation of bPRL (Bovine Prolactin) gene and its association to milk production in Indonesian Friesian Holstein (IFH) dairy cattle. In total 49 DNA from whole blood were extracted and followed with PCR-RFLP to distinguish the genotype difference between individual samples. The fragment of bPRL gene was amplified using pair of primer forward 5’-GAGTCCTTATGAGCTTGATTCTT-3’ and reverse 5’-GCCTTCCAGAAGTCGTTTGTTTTC-3’, resulting in 156 bp PCR product size. DNA digestion then performed using RsaI restriction enzyme that recognizes 5’-GT↓AC-3’ sites with blunt-end termini. According to the study, two genotypes were discovered that are AA and AB, with respective frequencies of 0.84 and 0.16. Based on the genotype grouping of milk production data, AB genotype yielded 4016.63±1344 kg while AA genotype yielded 3314.32±1153 Kg of milk per lactation. However, there is no statistical (P>0,05) difference between both genotypes on the amount of milk yielded. It is concluded that bPRL gene was not associated with milk production traits in the studied Indonesian Dairy Cattle population.

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Molecular screening of bovine β-casein (CSN2) A2 variant in Indonesian Holstein dairy cattle as attempt to produce digestive friendly milk

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/902/1/012060 ◽

2021 ◽

Vol 902 (1) ◽

pp. 012060

Author(s):

A M P Nuhriawangsa ◽

T Mulyani ◽

G Pambuko ◽

R Vanessa ◽

Purwadi ◽

...

Keyword(s):

Dairy Cattle ◽

Enzymatic Digestion ◽

Cattle Breeding ◽

Mutation Site ◽

Single Nucleotide ◽

Site Analysis ◽

Pcr Rflp ◽

Allele Variant ◽

Dairy Cattle Breeding ◽

Variant Analysis

Abstract Cow’s milk-intolerance is a digestive problem on people who not able to digest milk. This problem may relate to the variant of (3-casein (CSN2), especially A1, suggested due to (3-casomorphins (BCM-7) formation during enzymatic digestion, for that selecting cattle free BCM-7 become a concern to produce digestive friendly milk. This study aimed to differentiate A1 and A2 allele variant of CSN2 gene in selected population of Indonesian Holstein cattle. In total 70 cows DNA were collected, and fragment of CSN2 exon 7 which contain Single Nucleotide Polymorphism (SNP) rs43703011 and rs43703013 were amplified. Variant analysis was done by mutation site analysis using PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) with Mspl restriction enzyme and DNA sequence for confirmation. Result shows 2 allele variants of (3-casein that are B type, representing A1 family variant, and A2 in mutation site rs43703013C>G. We found A2 allele in the studied population is superior in frequency than A1 (0.916 vs. 0.084). Of that, 8.6% cattle were heterozygotes that is BA2 and 91.4% were homozygotes A2A2. Cattle which carry A1 allele variant should be excluded from dairy cattle breeding program for further milk production free of BCM-7.

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Epidemiological surveillance ofCampylobacter jejuniin chicken, dairy cattle and diarrhoea patients

Epidemiology and Infection ◽

10.1017/s0950268809002039 ◽

2009 ◽

Vol 137 (8) ◽

pp. 1111-1120 ◽

Cited By ~ 29

Author(s):

J. L. HUANG ◽

H. Y. XU ◽

G. Y. BAO ◽

X. H. ZHOU ◽

D. J. JI ◽

...

Keyword(s):

Dairy Cattle ◽

Campylobacter Jejuni ◽

Rflp Analysis ◽

Domestic Animals ◽

Epidemiological Surveillance ◽

Contamination Rate ◽

Genotype Distribution ◽

Isolation Rate ◽

Pcr Rflp ◽

Milk Cows

SUMMARYCampylobacter jejuniof different subtypes were identified in chicken, dairy cattle and diarrhoea patients in China from 2005 to 2006, using multiplex PCR and RFLP. The results indicated that, of the three types of samples,C. jejuniwas most frequently detected in poultry of the three types of samples, with an average isolation rate of up to 18·61% and a flock contamination rate of 86·67%. The average incidence ofC.jejuniin overall cattle and environmental samples, milk cows, heifers and diarrhoea patients was 7·77, 5·02, 8·70 and 4·84%, respectively. A higher prevalence was detected in outpatients than ward patients (P<0·01), and in patients aged <7 years than in older patients (P<0·01). The 265 isolates ofC.jejuniwere classified into 20 distinct types by PCR–RFLP analysis of theflaAgene, with the genotype distribution in humans overlapping that in poultry and cattle. This suggests that certainC. jejunistrains circulate between humans and domestic animals such as cattle and poultry.

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Effects of Retinoic Acid and 1,25-Dihydroxyvitamin D3 on IFN-gamma Secretion by Mononuclear Leukocytes from Nulliparous and Postparturient Dairy Cattle

International Journal for Vitamin and Nutrition Research ◽

10.1024/0300-9831.70.3.92 ◽

2000 ◽

Vol 70 (3) ◽

pp. 92-101 ◽

Cited By ~ 14

Author(s):

Burim Ametaj ◽

Brian Nonnecke ◽

Ronald Horst ◽

Donald Beitz

Keyword(s):

Retinoic Acid ◽

Dairy Cattle ◽

Dairy Cows ◽

Dairy Cow ◽

Pokeweed Mitogen ◽

Mononuclear Leukocytes ◽

Combined Effects ◽

Ifn Gamma ◽

Dihydroxyvitamin D ◽

1,25 Dihydroxyvitamin D

Individual and combined effects of several isomers of retinoic acid (RA) and 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) on interferon-gamma (IFN-gamma) secretion by blood mononuclear leukocytes (MNL) from nulliparous and postparturient Holstein cattle were evaluated in vitro. In the first experiment, effects on incubation period (24 to 72 hours) and time of supplementation (0 to 32 hours) with all-trans, 9-cis, 13-cis-, and 9,13-dicis-RAs (0 to 100 nM) on IFN-gamma secretion by pokeweed mitogen (PWM)-stimulated (0 and 10 mug/ml) MNL from nulliparous cattle were evaluated. In the second experiment, MNL from postparturient cows (bled at 0, 2, 4, and 16 days postpartum) were stimulated with PWM (0 and 10 mug/ml) in the presence of RA isomers (9-cis- or 9,13-dicis-RA; 0 to 100 nM), 1,25-(OH)2D3 (0 to 100 nM), or with combinations of these metabolites. The results show that individual isomers of RA had no effect on IFN-gamma secretion by PWM-stimulated MNL from nulliparous or postparturient cows. Furthermore 1,25-dihydroxyvitamin D3 inhibited IFN-gamma secretion by MNL from nulliparous and postparturient dairy cows; however, the degree of inhibition was greater when 9-cis- and 9,13-dicis-RA were also present in the cultures. Finally mononuclear leukocytes from postparturient dairy cows produced substantially less IFN-gamma than did MNL from nulliparous cattle. It is concluded that retinoic acids individually did not affect the capacity of leukocytes from dairy cattle to secrete IFN-gamma. This result is in marked contrast to studies in monogastric species indicating that RAs inhibit IFN-gamma secretion by peripheral blood T cells. Inhibition of IFN-gamma secretion by 1,25-(OH)2D3 was potentiated by 9-cis- and 9,13-di-cis-retinoics acids, suggesting that an excess of dietary vitamins A and D may compromise further the naturally immunosuppressed postparturient dairy cow. Additional research is necessary to determine if the combined effects of these metabolites on IFN-gamma secretion represent an increased susceptibility of the dairy cow to infectious diseases during the periparturient period. Lower secretion of IFN-gamma by MNL from postpartutient dairy cows, relative to nulliparous cattle, suggests that recently-calved cows are naturally immunosuppressed.

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