• Asian-Australasian Journal of Animal Sciences
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• 제8권2호
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• pp.187-194
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• 1995

Genetic variants of ${\alpha}s_1$-casein, ${\beta}$-casein, ${\kappa}$-casein and ${\beta}$-lactoglobulin were investigated by starch urea gel electrophoresis in milk samples of 280 Korean native cattle. A new ${\beta}$-casein variant, designated ${\beta}$-casein $A^4$, was found in milk samples of Korean native cattle. It has a much slower electrophoretic mobility than the ${\beta}$-casein $A^3$ variant in acid gel. This new variant appeared together with either ${\beta}$-casein $A^1$, $A^2$ or B variant. Gene frequencies and genotypic frequencies were estimated. Gene frequencies of four milk protein loci in Korean native cattle were compared with those of imported cattle breeds raised in Korea and Japanese brown cattle. Gene frequencies were ${\alpha}s_1$-casein B .846, ${\alpha}s_1$-casein C .154; ${\beta}$-casein $A^1$ .216, ${\beta}$-casein $A^2$ .666, ${\beta}$-casein $A^4$ .048, ${\beta}$-casein B .070; ${\kappa}$-casein A .648, ${\kappa}$-casein B .352; ${\beta}$-lactoglobulin A .148, ${\beta}$-lactoglobulin B .852. The population was in Hardy-Weinberg equilibrium at all milk protein loci. Gene frequencies of Korean native cattle were very similar to those of Japanese brown cattle. Interestingly, a new variant, ${\beta}$-casein $A^4$, was found only in Korean native cattle and Japanese brown cattle. These results support the hypothesis that Korean native cattle were used in the development of the Japanese brown cattle.

• 농업과학연구
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• 제25권2호
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• pp.216-224
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• 1998

본 연구는 축협중앙회 한우개량부에서 사육 중인 한우 암소 253두와 축산기술연구소에서 사육 중인 Holstein 113두의 혈액으로 부터 genomic DNA를 추출하고, PCR-RFLP기법에 의해 ${\beta}$-lactoglobulin과 ${\kappa}$-casein 유전자좌위의 유전적 다형을 분석하여 한우와 유우 집단에 대한 이들 유전자의 유전적 구조를 분석함으로써 한우와 유우 개량을 위한 기초 및 응용자료를 제공하고자 실시하였던 바 얻어진 결과를 요약하면 다음과 같다. 1. 한우와 Holstein종의 genomic DNA로 부터 PCR기법을 이용하여 ${\beta}$-lactoglobulin 과 ${\kappa}$-casem의 유전자좌를 증폭한 결과 각각 530bp와 262bp의 단편이 증폭되었음을 확인하였다. 2. ${\beta}$-lactoglobulin 증폭 산물에 대한 Hae III 제한효소의 처리결과, ${\beta}$-lactoglobulin AA형은 153bp와 109bp의 단편을, AB형은 153bp, 109bp, 79bp 및 74bp의 단편을, 그리고 BB형은 109bp, 79bp 및 74bp의 단편을 나타내었다. 3. ${\kappa}$-casein 유전자좌의 증폭산물에 대한 Taq I의 제한효소 처리결과, ${\kappa}$-casein AA형은 530bp의 단편을, AB형은 530bp, 344bp 및 186b의 단편을, 그리고 BB형은 344bp 및 186mp의 단편을 나타내었다. 4. ${\beta}$-lactoglobulin의 유전자형 및 유전자 빈도에 있어서, 한우의 ${\beta}$-lactoglobulin AA, AB 및 BB 유전자형 의 빈도는 각각 6.72, 26.09 및 67.19%이었으며, ${\beta}$-lactoglobulin A 및 B 유전자 빈도는 각각 0.197 및 0.803이었고, Holstein종의 ${\beta}$-lactoglobulin AA, AB 및 BB 유전자형 빈도는 각각 35.40, 56.64 및 7.96%이었으며, ${\beta}$-lactoglobulin A 및 B 유전자 빈도는 각각 0.637 및 0.363이었다. 5. ${\kappa}$-casein의 유전자형 및 유전자 빈도에 있어서, 한우의 ${\kappa}$-casein AA, AB 및 BB 유전자형의 빈도는 각각 46.25, 39.13 및 14.62%이었으며, K-casein A 및 B 유전자 빈도는 각각 0.658 및 0.342이었고, Holstein종의 ${\kappa}$-casein AA, AB 및 BB유전자형 빈도는 각각 60.18, 38.94 및 0.88%이었고, ${\kappa}$-casein A 및 B 유전자 빈도는 각각 0.796 및 0.204 이었다. 6. 이상의 결과로써 ${\beta}$-lactoglobulin과 ${\kappa}$-casein의 유전자 빈도는 한우에서 ${\beta}$-lactoglobulin A 및 B 대립 유전자 빈도는 각각 0.197 및 0.803이었고 ${\kappa}$-casein A 및 B 대립유전자 빈도는 각각 0.658 및 0.342이었다. 그러나 Holstein 종에서는 ${\beta}$-lactoglobulin A 및 B 대립 유전자 빈도는 0.637 및 0.363이었고, ${\kappa}$-casein A 및 B 대립유전자 빈도는 각각 0.796 및 0.204이었다.

• Asian-Australasian Journal of Animal Sciences
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• 제11권3호
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• pp.311-317
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• 1998

A total of 1033 Brown Swiss and 610 Canadienne cows were phenotyped for the genetic variants ${\alpha}_{s1}$-casein, ${\beta}$-casein, ${\kappa}$-casein, ${\beta}$-lactoglobulin and ${\alpha}$-lactalbumin. In Brown Swiss, frequency distributions were: 97.3% B and 2.7% C variant of ${\alpha}_{s1}$-casein; 31.6% $A^1$, 51.8% $A^2$, 0.5% $A^3$ and 16.1% B variant of ${\beta}$-casein; 70.4% A, 29.3% B, and 0.3% C variant of ${\kappa}$-casein; 41.7% A and 58.3% B variant of ${\beta}$-lactoglobulin; and 100% B variant of ${\alpha}$-lactalbumin. Corresponding frequencies in Canadienne for those five milk proteins were: 98.6 and 1.4%;58.5, 33.5, 0.08 and 7.9%; 78.8, 21.1 and 0.1%, 42.4 and 57.6%; and 100%. Analysis of variance by least squares showed possible association between milk protein phenotypes and some lactational production traits. There were no significant association of phenotypes of ${\alpha}_{s1}$-casein, ${\beta}$-casein and ${\beta}$-lactoglobulin with milk yield, fat yield, protein yield, fat percentage and protein percentage in both breeds during the three lactations. In the Brown Swiss, ${\kappa}$-casein phenotype was associated with 305-day fat yield and protein yield during the first lactation. ${\kappa}$-Casein AB was associated with higher milk, fat and protein yield during the second lactation. During the third lactation, ${\beta}$-lactoglobulin AA in Canadienne cows was associated with higher protein content in the milk (3.70%) when compared to phenotypes AB (3.54%) and BB (3.64%).

• Reproductive and Developmental Biology
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• 제41권3호
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• pp.51-55
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• 2017

The production of therapeutic protein from transgenic domestic animal is the major technology of biotechnology. Insulin-like growth factor-1 (IGF-1) is known to play an important role in the growth of the animal. The objective of this study is construction of knock-in vector that bovine IGF-1 gene is inserted into the exon 7 locus of ${\beta}$-casein gene and expressed using the gene regulatory DNA sequence of bovine ${\beta}$-casein gene. The knock-in vector consists of 5' arm region (1.02 kb), bIGF-1 cDNA, CMV-EGFP, and 3' arm region (1.81 kb). To express bIGF-1 gene as transgene, the F2A sequence was fused to the 5' terminal of bIGF-1 gene and inserted into exon 7 of the ${\beta}$-casein gene. As a result, the knock-in vector is confirmed that the amino acids are synthesized without termination from the ${\beta}$-casein exon 7 region to the bIGF-1 gene by DNA sequence. These knock-in vectors may help to create transgenic dairy cattle expressing bovine bIGF-1 protein in the mammary gland via the expression system of the bovine ${\beta}$-casein gene.

• 한국가축번식학회지
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• 제22권3호
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• pp.237-244
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• 1998

To investigate the fidelity of transgene transmission and expression, we produced transgenic mice carrying bovine $\beta$-casein/bovine grwoth hormone(bGH) fusion gene and examined transmission efficiency and expression level of the transgene in the founders and their progeny. The transgene was composed of 1.8 kb bovine $\beta$-casein promoter and 2.1 kb bGH gene. Ten transgenic mice were produced. Milk and mammary gland were collected from eight transgenic lines at 10-day lactation and a, pp.ied to Western and Northern blot analyses. The bGH expression was detected in four of them. The concentrations of bGH in milk were highly variable from 4$\mu\textrm{g}$/ml to 600$\mu\textrm{g}$/ml depending on the lines. The bGH mRNA level in mammary gland was closely correlated with the bGH concentration in milk in each transgenic line. These results indicated that bGH transgene expression was a, pp.opriately regulated in the mammary gland and secreted into milk in transgenic mice. By using two transgenic lines(#2, #7) secreting a considerable amoung of bGH into their milk, the inferitance and maintenance of transgenic phenotype were assessed in successive four generations. The mean transmission frequencies of transgene in lines #2 and #7 were 34% and 40%, respectively. The bGH concentration in milk were 80, 240, 120, 60$\mu\textrm{g}$/ml in each G0(generation 0), G2, G3, G4 generation of line #2 and 600, 1600, 860, 900$\mu\textrm{g}$/ml in each G1. G2, G3, G4 generation of line #7. These results demonstrated that bovine $\beta$-casein/bGH gene was stably transmitted from generation to generation in a Menelian fashion in trasgenic mice and consistenly expressed in their milk throughout the generations, although there was a little variation in the transmission frequency and expression level of the transgene between generations.

• 농업과학연구
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• 제29권2호
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• pp.43-52
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• 2002

본 연구는 한국 재래 산양 112두와 유산양인 Saanen종 7두의 혈액으로부터 genomic DNA를 추출하고, PCR-RFLP 방법에 의해 $\beta$-casein 유전자의 특성을 분석하여 한국재래산양의 효율적인 유전자원의 보존 및 개량을 위한 기초 자료로 제공하고자 실시하였다. 한국재래산양의 genomic DNA로부터 PCR기법을 이용하여 $\beta$-casein의 유전자좌를 증폭한 결과 각각 481bp 크기의 단편이 양호하게 증폭되었음을 확인하였다. $\beta$-casein 유전자 좌의 증폭산물에 대한 Bal I의 제한효소를 처리한 결과, $\beta$-casein AB형은 481bp, 284bp 및 197bp의 단편을, 그리고 BB형은 284bp와 197bp의 단편을 한국재래산양과 유산양인 Saanen 종에서 확인 할 수 있었다. 유전자형 빈도에 있어서는 한국재래산양에서 $\beta$-casein AB 및 BB의 빈도는 각각 6.25 및 93.75%이었고, 유산양인 Saanen 종은 각각 57.14 및 42.86%이었다. 유전자빈도에 있어서는 한국재래산양의 $\beta$-casein A 및 B의 빈도가 각각 0.031 및 0.969이었고, Saanen 종에서는 각각 0.286 및 0.714의 빈도를 보였다. 한국재래산양의 $\beta$-casein 유전자의 염기서열과 이미 보고되어 있는 goat의 염기서열(GeneBank accession Number M90556)간에는 총 11개의 염기서열에 차이를 나타내어 97.71%의 상동성을 보였다. 따라서 한국재래산양의 $\beta$-casein 유전자의 다형성과 염기서열 분석에 의한 분자유전학적 특성의 규명은 한국재래산양의 유전자원의 보존 및 개량을 위한 기초 및 응용 자료로 이용될 수 있을 것으로 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제16권8호
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• pp.1212-1217
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• 2003

Fifty-two Holstein cows with different phenotypes of $\kappa$-casein ($\kappa$-CN) and $\beta$-lactoglobulin ($\beta$-LG) were selected to provide weekly milk samples for heating at 30, 70, 75 and $80^{\circ}C$ for 2 min. Coagulating properties of heated milk samples measured as rennet clotting time, rate of curd firming and curd firmness at cutting were determined by a Formagraph. Milk samples were analysed for fat and casein. Least squares analyses of data, after adjustments were made for effect of milk casein and fat contents, indicated that although an increase in heating temperatures resulted in less desirable coagulating properties, the effect of milk types was inherent irrespective of heating temperatures. The shortest rennet clotting time (6.06 min), fastest rate of curd firming (5.61 min) and firmest curd (38.05 mm) were obtained from milk with the B variant for $\kappa$-CN and B variant for $\beta$ -LG when preheated at $30^{\circ}C$. It appears that milk bearing $\kappa$-CN B is more resistant to heat perturbation. All milk samples having the $\kappa$-casein AA (milk types AA/AA, AA/AB, AA/BB) did not have a measurable K20 value when preheated at $70^{\circ}C$. This effect was observed for $\kappa$-casein AB (milk types AB/AA, AB/AB, AB/BB) at $75^{\circ}C$ and $\kappa$-casein BB (milk types BB/AA, BB/AB, BB/BB) at $80^{\circ}C$.

• Journal of Dairy Science and Biotechnology
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• 제25권2호
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• pp.11-16
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• 2007

The nomenclature of genetic variants of casein which is major protein in milk have had a lot of confusion, but now have established. Genetic variants of ${\alpha}_{s1}-CN,\;{\alpha}_{s2}-CN,\;{\beta}-CN,\;{\kappa}-CN$ have reported 8 variants(A, B, C, D, E, F, G, H), 4 variants(A, B, C, D), 13 variants ($A_1,\;A_2,\;A_3,\;A_4$, B, C, D, E, F, G, $H_1,\;H_2$, I), 11 variants(A, B, C, E, $F_1,\;F_2,\;G_1,\;G_2$, H, I, J), respectively. Their data detailed have introduced in several web sites including www.uniprot.org. The studies on genetic variants of casein from Korean native cattle have been reported only ${\beta}-casein\;A_4$ but still not established the protein sequence. The classification and distinct nomenclature of genetic variants of bovine casein were required because the development of milk science and technology have been focused in the region that have to studied biochemically such as functional foods, EMC and GMO et al.

• Asian-Australasian Journal of Animal Sciences
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• 제9권6호
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• pp.685-693
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• 1996

A total of 3,610 Ayrshire and 1,711 Jersey cows were phenotyped for the genetic variants of ${\alpha}_{s1}$-casein, ${\beta}$-casein, $\chi$-casein, ${\beta}$-lactoglobulin and ${\alpha}$-lactalbumin. Least squares analyses showed possible associations between milk protein phenotypes and lactational production traits. Depending on lactation number, ${\beta}$-casein phenotypes in Ayrshires were associated with milk production ($A^2A^2$ > $A^1A^2$ > $A^1A^1$), and with milk protein content. In the third lactation, Ayrshire cows with ${\beta}$-casein $A^1A^1$ produced milk with 3.43% fat compared to 3.37% fat for ${\beta}$-casein $A^2A^2$. In Ayrshire, $\chi$-casein phenotypes affected the protein content during the three lactations (BB > AB > AA) and ${\beta}$-lactoglobulin phenotypes significantly influenced the milk fat during the first lactation (4.06% for AA and 3.97% for BB). In Jerseys, protein content of milk was influenced by phenotypes of ${\alpha}_{s1}$-casein(3.98% for CC v/s 3.86% for BB in the first lactation). In the third lactation, $\chi$-casein AA of Jersey milk contained 5.35% fat compared to 4.82% for phenotype BB. The effects of ${\beta}$-lactoglobulin phenotypes on protein content were apparent in Jerseys during the second lactation with the A variant being superior to the B (4.00% for AA v/s 3.87% for BB).

• Asian-Australasian Journal of Animal Sciences
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• 제32권1호
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• pp.14-22
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• 2019

Objective: The aim of the study was to find a possible association between the ${\beta}-$ and ${\kappa}-casein$ and ${\beta}-lactoglobulin$ genotypes and important milk physiochemical and technological characteristics such as acidity, alcohol stability, the contents of some minerals and the parameters of acid fermentation ability (FEA) in Czech Fleckvieh Cattle. Methods: Milk and blood samples were collected from 338 primiparous Czech Fleckvieh cows at the same stage of lactation. The genotypes of individual cows for ${\kappa}-casein$ (alleles A, B, and E) and ${\beta}-lactoglobulin$ (alleles A and B) were ascertained by polymerase chain reaction-restriction fragment length polymorphism, while their ${\beta}-casein$ (alleles $A^1$, $A^2$, $A^3$, and B) genotype was determined using melting curve genotyping analysis. The data collected were i) milk traits including active acidity (pH), titratable acidity (TA), alcohol stability (AS); calcium (Ca), phosphorus (P), sodium (Na), magnesium (Mg), and potassium (K) contents; and ii) yoghurt traits including active acidity (Y-pH), titratable acidity (Y-TA), and the counts of both Lactobacilli and Streptococci in 1 mL of yoghurt. A linear model was assumed with fixed effects of herd, year, and season of calving, an effect of the age of the cow at first calving and effects of the casein and lactoglobulin genotypes of ${\beta}-CN$ (${\beta}-casein$, CSN2), ${\kappa}-CN$ (${\kappa}-casein$, CSN3), and ${\beta}-LG$ (${\beta}-lactoglobulin$, LGB), or the three-way interaction between those genes. Results: The genetic polymorphisms were related to the milk TA, AS, content of P and Ca, Y-pH and Lactobacilli number in the fresh yoghurt. The CSN3 genotype was significantly associated with milk AS (p<0.05). The effect of the composite CSN2-CSN3-LGB genotype on the investigated traits mostly reflected the effects of the individual genes. It significantly influenced TA (p<0.01), Y-pH (p<0.05) and the log of the Lactobacilli count (p<0.05). Conclusion: Our findings indicate that the yoghurt fermentation test together with milk proteins genotyping could contribute to milk quality control and highlight new perspectives in dairy cattle selection.