• Title/Summary/Keyword: casein

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Expression of Human Serum Albumin in Milk of Transgenic Mice Using Goat β-casein/Human Serum Albumin Fusion Gene

  • Wu, H.T.;Chou, C.K.;Huang, M.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.743-749
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    • 2004
  • The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.

Isolation and Characteristics of Lactic Acid Bacteria Producing Casein Phosphopeptides from Kimchi (Casein phosphopeptide를 생산하는 김치 유래 유산균의 분리 및 특성 연구)

  • Lee, Mi-Gyeong;Kwon, Hyo-Jeong;Byun, Ock-Hee;Bang, Bo-Yeon;Kim, Yu-Jin;Park, JungMin;Bai, Dong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.68-73
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    • 2016
  • Lactic acid bacteria showing both protease activity and the capacity to produce casein phosphopeptide (CPP) were isolated from Korean kimchi, a traditional food made from fermented vegetables. Among the 450 strains of isolated lactic acid bacteria, the strain MG-379 showed high protease activity and the highest ability to produce CPP. Characterization results showed that MG-379 was gram-positive and measured $0.6-0.8{\mu}m$ in diameter. DNA sequencing of MG-379 and comparison with other sequences using BLAST revealed a 100% identity with the sequence of Enterococcus faecalis. However, MG-379 showed a higher CPP-producing ability than E. faecalis KCCM 40450. Accordingly, MG-379 was newly named as E. faecalis MG-379. Amount of free calcium liberated by CPPs was 2227.5 and 1151.6 mg/kg for E. faecalis MG-379 and E. faecalis KCCM (control), respectively.

The Expression Changes of Casein mRNAs in Mammary Epithelial Cells Recovered from Bovine Milk during the Lactation Period

  • Ishii, Hiroshi;Nakamura, Tadashi;Higuchi, Munenori;Mamada, Aya;Fukushima, Michihiro;Urashima, Tadasu;Arai, Ikichi
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.983-988
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    • 2007
  • The aim of this study was to examine the correlation between bovine casein (CN) mRNA expression levels in mammary epithelial cells and lactation period, the yields of milk proteins and other parameters. The cells were collected from each cow's milk, which contained somatic cell counts (SCC) of less than 100,000 cells/ml. The levels of ${\alpha}s1-$, ${\alpha}s2-$, ${\beta}$- and ${\kappa}$-CN mRNA expression were significantly correlated with each other in mammary epithelial cells (p<0.01). All cows produced either less than 30 kg/day/cow or a over 30 kg/day/cow level of milk yield (MY). It was shown that the CN mRNA expression levels decreased gradually from the calving period to late lactation, when MY was over 30 kg/day/cow. The SCC tended to increase gradually during the course of lactation, but it was negatively correlated with milk protein and CN yields (p<0.01) when MY was less than 30 kg/day/cow. Moreover, there was a tendency for a negative correlation between SCC and ${\alpha}s1$-CN and ${\beta}$-CN mRNA expression level, when MY was less than 30 kg/day/cow (p<0.05).

Effects of Soy Protein and Isoflavones on Bone Markers and Hormones in Growing Male Rats (콩 단백질과 Isoflavones가 성장기 수컷 흰쥐에서 골 지표와 호르몬에 미치는 영향)

  • 최미자
    • Journal of Nutrition and Health
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    • v.36 no.5
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    • pp.452-458
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    • 2003
  • Soybean is a rich source of isoflavones such as genistein and daidzein. Soy isoflavones have both weak estrogenic and anti-estrogenic effects and are structurally similar to tamoxifen, an agent that has an effect similar to that of estrogen in terms of reducing postmenopausal bone loss. The purpose of this study was to determine the effects of differences in protein source (casein vs soy) and isoflavone levels (reduced vs higher levels) on selected bone markers and hormones in growing male rats. Thirty weanling Sprague-Dawley young rats were divided into 3 groups: The control group was fed a casein-based diet, the soy concentrate group was fed soy protein with totally reduced isoflavones content (isoflavones 0.07 mg/g protein), and the soy isolate group was fed soy protein with a higher than normal isoflavones content (isoflavones 3.4 mg/g protein). The degree of bone formation was estimated by measuring serum osteocalcin and alkaline phosphoatase (ALP). By determining collagen cross-linkage by immunoassay and correcting with creatinine values, the bone resorption rate was compared. Serum osteocalcin, growth hormone, estrogen and calcitonin were analyzed using radio immunoassay kits. The bone formation marker and ALP activity were differentiated by protein source, showing higher values than casein in feeding either soy isolate or soy concentrate. In this study using growing rats, the differences in isoflavone contents were not a significant factor in either bone formation or bone reaborption markers. Moreover, the soy isolate group had significantly higher levels of growth hormone than the casein group. The findings of this study suggest that growth hormone is partially responsible for its bone-formation effects in young growing rats. Soy protein and the isoflavones in soy protein are beneficial for bone-formation in growing male rats. Therefore, exposure to soy protein and isoflavones early in life may have long-term health benefits in preventing bone diseases such as osteoporosis. Further study to evaluate the mechanism of action of isoflavones on bones is warranted. (Korean J Nutrition 36(5): 452∼458, 2003)

Effect of the Toluene Administration on the Liver Damage in Rats Fed Low or Standard Protein Diet (저 및 표준단백식이로 성장시킨 흰쥐에 Toluene 투여가 간 손상에 미치는 영향)

  • 윤종국;김경순;임영숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.6
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    • pp.981-985
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    • 1996
  • To study the effect of toluene administration on the liver damage, rats were previously fed a low (casein 7%, LP) or standard(casein 20%, SP) protein diet and for four days toluene(50% in olive oil) was given at 0.2ml/100g body weight/day to the male rats, and then the degree of liver damage in toluenetreated animals fed LP were compared with those fed SP. The increasing rate of liver weight/body weight and the serum levels of xanthine oxidase to the control group were higher in rats fed SP than those fed LP. The decreasing rate of protein contents in cytosol, mitochondria and glycogen, glutathione contents of liver to the control group were higher in rats fed SP than those fed LP. In histopathological findings, the swelling of hepatic cell around the central vein was demonstrated in all the two groups toluene-treated rats. But the degree of swelling severity in hepatocytes was somewhat higher in rats fed SP than those fed LP. Therefore it is assumed that the degree of liver damage severity in toluenetreated animals was higher in rats fed SP than those fed LP.

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A Study of the Free Amino Acids in the Plasma and Erythrocytes in the Male Adult Rats Fed with the Low Protein Diets (저단백영양(低蛋白營養)에 있어서 성숙(成熟)흰쥐의 Plasma 및 Erythrocytes Free Amino Acid에 대하여)

  • Lee, Hyun-Ki
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.1 no.1
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    • pp.37-50
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    • 1972
  • An analysis of the free amino acid contained in the plasma and erythrocytes of the six groups of Wistar Strain male adult rats(body weight 200-300g) having fasted for sixteen hours was made by means of the HITACHI Amino Acid Autoanalyzer and the result of which was corrected with RC-24 B TOMY Micro Hematocrit Centrifuge. There was a depression of the plasma and erythrocytes free amino acid level on the noprotein diet with ad libitum feeding. But on the 20% casein diet there was an elevation in the levels of free amino acid and consequently alanine, glysine, lysine, serine and arginine level in the erythrocytes and threonine glutamic acid and taurine level in the plasma increased on the high protein diet. There was more plasma and erythrocytes free amino acid level on the 5% casein-30% fat diet than on the 5% casein-no fat diet with pair-feeding. In comparison, on the low calorie diet more free amino acids were found in plasma than in erythrocytes, but on the higher calorie diet more free amino acids were found in the erythrocytes than in the plasma. On the 20% casein-30% fat diet with pair-feeding the erythrocytes free amino acids level increased but in plasma free amino acids level decreased. Such as an opposite result was given in plasma and erythrocytes free amino acids level. In the pair-fed four groups, erythrocytes per plasma generally increased in the rate of less than 10.0 as the calorie increased. The essential amino acid per non essential amino acid generally increased in the ratio as protein level and calorie increased, and that ratio range was from 0.2 to 0.7. And essential amino acid per non essential amino acid of plasma was higher than that of erythrocytes.

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Heterologous Introns Enhanced Expression of Human Lactoferrin cDNA in Mouse Mammary Epithelial Cells

  • Kim, Sun-Jung;Yu, Dae-Yeul;Lee, Ko-Woon;Cho, Yong-Yeon;Lee, Chul-Sang;Han, Yong-Mahn;Lee, Kyung-Kwang
    • BMB Reports
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    • v.28 no.1
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    • pp.57-61
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    • 1995
  • The expression of a recombinant human lactoferrin is reported in mouse HC11 mammary epithelial cells. Expression of human lactoferrin (hLF) was achieved by placing its cDNA under the control of the bovine ${\beta}$-casein gene. To improve the hLF expression level in a cell culture system, two artificial introns were also introduced to construct expression vectors. One intron was a hybrid-splice signal consisting of bovine ${\beta}$-casein intron 1 and rabbit ${\beta}$-globin intron II. The other intron was a DNA fragment spanning intron 8 of the bovine ${\beta}$-casein gene. The hybrid intron moderately elevated hLF expression, whereas intron 8 alone did not express any detectable amount of hLF as judged by Northem and Western blot analyses. When the two introns were used together they contributed to a synergistic elevation of hLF expression. These data indicate that artificial introns on both sides of the hLF cDNA were necessary to increase expression of cDNA.

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Analysis of Changes in Colostrum Proteins by Mammalian Species (포유류의 종에 따른 초유 단백질의 변화에 대한 분석)

  • Kim, Seung Hee;Kim, Woan-Sub
    • Journal of Dairy Science and Biotechnology
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    • v.35 no.2
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    • pp.105-111
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    • 2017
  • There have been numerous reports indicating that milk proteins influence immune functions. Colostrum refers to the breast milk of mammals, secreted starting from the fourth or fifth day after delivery. It has abundant nutrition for the survival of newborn infants. Most importantly, it contains bioactive substances with growth-stimulating and antibiotic, functions. Thus, the colostrum has various physiological roles. This study measured the differences in the composition of colostrum derived from dairy cattle, hanwoo, porcine, and goat sources. The results showed that immunoglobulin, lactoferrin, lactoperoxidase, serum albumin, IgG heavy chain, and IgG light chain were significantly higher in the colostrum of dairy cattle, hanwoo, and goats, but low in porcine colostrum. There was no significant difference in ${\alpha}_{S2}$-casein, ${\alpha}_{S1}$-casein, ${\beta}$-casein, ${\kappa}$-casein, ${\beta}$-lactoglobulin, and ${\alpha}$-lactalbumin contents until seven days after birth. However, porcine colostrum showed high contents of all proteins from the first day to the second day after delivery.

Protein Qualify of Noodles Supplemented with Fish Protein Concentrate (농축(濃縮) 생선단백질(生鮮蛋白質)로 보충(補充)된 국수의 단백질(蛋白質)의 질(質)에 대(對)한 연구(硏究))

  • Choi, Hay-Mie
    • Journal of Nutrition and Health
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    • v.9 no.1
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    • pp.99-103
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    • 1976
  • 농축생선단백질(Fish Protein Concentrate, FPC)로 보충된 국수의 영양가를 연구하기 위하여 25일된 쥐를 사용하여 4주 동안 동물 실험을 하였다. 각각 10마리씩 네 group으로 나누어 0, 10, 15% FPC와 표준 casein diets를 주고 매일 섭취량을 기록하고, 매주 동물의 체중을 재서 Protein Efficiency Ratio (PER)를 계산함으로써 단백질의 질(質)을 평가하였다. 4주 후에 동물의 간을 취하여 수분과 지방의 양을 측정하였다. 또한 농축생선 단백질로 보충된 국수가 음식으로서의 적응도를 측정하기 위하여 어른 10명과 아이들 5명에게 관능 검사를 실시하였다. FPC 15% diet group의 체중은 4주 동안에 157.2g이 증가했고, FPC 10%는 135.7g, casein은 103.0g, FPC 0%(plain noodle)은 25.8g 밖에 되지 않았다. 이것들의 PER은 FPC 15% diet group 이 3.83으로 제일 높았고, 다음이 FPC 10%로 3.35였고, casein은 이보다 낮은 3.31이였다. 간의 지방양은 FPC 15% diet group이 2.94%로 제일 낮았으며, FPC 10%가 3.98%, casein이 5.57%의 순서였다. 지방의 양이 적을수록 단백질의 질(質)이 높다는 것을 말하며, 5%정도가 보통이다. PER, 동물의 체중과 지방의 양으로 단백질의 질(質)을 측정한 결과 FPC 15% 국수가 질(質)이 제일 좋다는 것이 판명되었다. 관능검사 결과 전체적 계수는 FPC 15% 국수가 제일 낮은 2.7 (1.0에서 5.0까지 사용)이고 다음이 FPC 10% 국수로 3.5였으나 plain noodle만 못하였다. 특히 색갈(color)에 많은 차이를 보였기 때문이며, 맛(flavor)과 질(texture)에는 별 차이가 없었다. FPC 15% 국수는 색갈이 plain noodle에 비해 검은 편이였으나 모밀국수 보다는 색갈이 희므로 그렇게 문제가 될것 같지는 않다. 어린 아이들에게는 닭 국물에 국수를 넣어 주였더니 라면과 맛이 비슷해 아주 좋아했다.

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Improving the Surface Functionality of Curdlan by Conjugation with Unfolding Protein through Naturally Occurring Maillard Reaction

  • Nakamura, Soichiro;Ogawa, Masahiro;Saeki, Hiroki;Saito, Masayoshi;Miyasaka, Satoko;Hata, Junya;Adachi, Naoko;Hwang, Jae-Kwan
    • Preventive Nutrition and Food Science
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    • v.5 no.4
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    • pp.200-204
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    • 2000
  • Protein conjugation of curdlan belonging to $\beta$-1, 3-glucan was carried out to improve it surface functionalities. The glucan was mixed with phosvitin, {TEX}$$\alpha$_{s}${/TEX}-casein, lysozyme or ovalbumin, respectively. The mixture was freeze-dried, and he resulting powder was incubated at 6$0^{\circ}C$ and 79% relative humidity for 12 days in order to generate a controlled Maillard reaction between curdlan and proteins. conjugation with unfolding proteins, i.e., phosvitin and {TEX}$$\alpha$_{s}${/TEX}-casein, drastically increased the solubility of the glucan, whereas lysozyme and ovalbmin did not. The solubility in water of curdlan was 3.44% for the phosvitin conjugate and 1.09% for the {TEX}$$\alpha$_{s}${/TEX}-casein conjugate. SDS-slab polyacrylamide gel electrophoresis showed that curdlan was solubilized due to covalent binding with phosvitin. Emulsifying properties of curdlan were substantially improved by the conjugation with phosvitin and {TEX}$$\alpha$_{s}${/TEX}-casein. Emulsion stability of the curdlan-phosvitin conjugate was about 2.9 times greater than that of the curdlan-phosvitin mixture.

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