• Journal of Animal Science and Technology
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• 제44권6호
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• pp.801-808
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• 2002

상업적 단백질 분해 효소에 0.02% $CaCl_2$를 첨가하여 응유 활성화를 시킨 탈지유에 대한 분해 작용의 결과를 요약하면 다음과 같다. 다양한 효소별 가수분해 시간에 따른 가수분해도는 미생물 유래 효소와 trypsin은 pepsin과 papain W-40보다 높은 분해도를 나타냈다. 12% TCA 용액에 가용성인 NPN의 양은 trypsin이 가장 높은 분해도를 나타내었고 rennet과 pepsin이 가장 낮은 분해도를 보였다. 전기영동에 있어서 trypsin과 protease S는 $\alpha$- lactalbumin을 분해하였고 papain w-40은 $\beta$- lactoglobulin을 미약하게 분해하였으며 neutrase 1.5는 90분 이후부터 $\alpha$-lactalbumin과 $\beta$-lactoglobulin을 분해하였다. Rennet과 비교한 전기영동상에서는 rennet에 의해 분해 되지 않은 ${\alpha}_s$- casein과 $\beta$-casein을 trypsin과 protease S가 다량 분해하였고 $\kappa$-casein은 rennet에 비해 papain W-40이 상당 수준의 분해상을 나타내었다. 이상의 결과 가수분해도 및 NPN 양은 trypsin, neutrase 1.5 및 protease S가 다른 효소에 비해 높게 나타났으며, 전기영동상에서는 pepsin과 neutrase 1.5가 rennet과 유사한 경향을 나타내었다.

• BMB Reports
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• 제31권4호
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• pp.413-417
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• 1998

Transgenic mice containing a bovine ${\beta}-Casein/Human$ lysozyme fusion gene (pBZ) were generated in order to produce human lysozyme in their milk. The expression vector was a quadripartite fusion consisting of a 2 kb upstream DNA of the bovine ${\beta}-casein$ gene, human lysozyme gene, intron II of the rabbit ${\beta}-globin$ gene, and the polyadenylation/termination signals of SV40 DNA. Fertilized mouse zygotes were microinjected with pBZ, then transferred into the oviduct of foster mothers. Out of 20 mice born, 11 survived until postweaning and three were identified as positivetransgenic by Southern blot analysis (one male and two females). The founder mice were mated to BCFl mice to produce transgenic progeny. It was confirmed by RT-PCR and Northern blot analyses that the transgene was specifically expressed in the mammary gland of the founder mice. Furthermore, the artificial introns within the transgenic RNA was proven to be correctly spliced out as judged by RT-PCR analysis. These results indicated that transgenic mice generated in this study properly expressed the human lysozyme RNA in their mammary gland.

• BMB Reports
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• 제28권1호
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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.

• 농업과학연구
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• 제28권2호
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• pp.92-98
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• 2001

락토페리신은 다양한 생리활성을 나타내는 락토페린(약 80kD)에서 유래된 항균성펩타이드 분획물(5kD)이다. 마우스HC11유선상피세포에서 인체 락토페리신의 발현은 bovine beta-casein을 promotor로 하고 인체 락토페리신 cDNA를 삽입하여 제작한 pBL1-cin발현벡타를 이용하였다. 이 발현벡타를 이용하여 인체 락토페리신 발현여부를 RT-PCR, northern blot, dot blot분석을 통하여 확인하였다. pBL1-cin 발현백타를 HC11세포에 transfection 하여 얻은 RNA를 이용하여 RT-PCR를 한 결과 150bp의 크기로 확인되었고 Northern blot 분석결과는 약 2.3 kb의 크기로 확인되었다. 인체 락토페린 polyclonal항체를 이용하여 dot blot한 결과 인체 락토페리신이 분비됨을 확인하였다.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2003년도 제3회 발생공학 국제심포지움 및 학술대회
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• pp.89-89
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• 2003

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2004년도 제4회 발생공학 국제심포지움 및 학술대회
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• pp.45-45
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• 2004

• Asian-Australasian Journal of Animal Sciences
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• 제6권4호
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• pp.541-547
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• 1993

A phage clone containing the partial ${\alpha}_{S1}$-casein gene was isolated from a bovine genomic library by using mixed probes of ovine ${\alpha}_{S1}$-, ${\beta}$- and ${\kappa}$-casein cDNAs. Restriction enzyme mapping analysis for 14.6 kb revealed that the map was in conflict with the report of Meade et al. (1990), especially in the 3'-end fragment. Sequence analysis of 12.6 kb revealed a high AT/GC ratio (1.64); we have identified eight exon sequences according to the bovine ${\alpha}_{S1}$-casein cDNA sequence. The same exon/intron splice junction sequence was observed between these exons. We suggest that the bovine ${\alpha}_{S1}$-casein gene night contain a minimum of 18 exons and the full length is approximately 18-19 kb.

• 농업과학연구
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• 제16권2호
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• pp.201-211
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• 1989

Mucor rennet의 이용(利用) 가능성(可能性)을 타진(打診)하기 위하여 Blue cheese를 제조(製造)하고 그 각종(各種) 질소화합물(窒素化合物)의 특성(特性)을 조사(調査)한 결과(結果)를 요약(要約)하면 다음과 같다. 1. Blue cheese의 수율(收率)은 calf rennet과 Mucor rennet간(間)에는 큰 차이(差異)가 없었다. 2. 숙성과정중(熟成過程中) cheese의 고형분(固形分) 함량(含量)은 증가(增加)하였고 첨가비율(添加比率)에 따른 차이(差異)는 인정(認定)되지 않았다. 3. water soluble nitrogen은 숙성(熟成)이 진행(進行)됨에 따라 증가(增加)하였으며 숙성초일(熟成初日)부터 calf rennet에 비(比)하여 Mucor rennet 편이 높았고 숙성(熟成) 40일(日)부터는 혼합(混合)rennet이 calf rennet보다 낮았다. 4. non protein nitrogen, peptone amino nitrogen, water soluble protein nitrogen, proteose nitrogen, peptone nitrogen 및 숙성율등(熟成率等)도 water soluble nitrogen과 동등(同等)한 변화(變化)를 보였다. 5. Mucor rennet으로 제조(製造)한 Blue cheese casein을 전기영동(電氣泳動)한 결과(結果) as-casein과 ${\beta}$-casein의 분해도(分解度)가 높았으나 calf rennet로 제조(製造)한 Blue cheese의 as-casein과 ${\beta}$-casein의 분해도(分解度)는 비교적 낮았고 숙성(熟成) 60일(日)에는 Mucor rennet 처리구(處理區)의 casein은 다소 잔존(殘存)함을 알 수 있었다. 6. Blue cheese를 60일간(日間) 숙성(熟成)시켰을때 유리(遊離)아미노산(酸) 함량(含量)은 Mucor rennet으로 제조(製造)한 cheese보다 전(全) 숙성기간(熟成期間)동안 많았다.

• Journal of Dairy Science and Biotechnology
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• 제35권2호
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• pp.105-111
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• 2017

젖소, 한우, 돼지, 그리고 산양의 초유 중 단백질의 변화를 초일부터 7일까지 조사하였다. 돼지의 초유를 제외한 젖소, 한우, 그리고 산양의 초유에서 면역글로불린, 락토페린, 락토퍼옥시데이스, 혈청 알부민, IgG heavy chain, 그리고 IgG light chain은 분만 후, 초일 함량이 현저히 높았고, 2일째부터 급격히 감소하는 것을 보여 주었다. 그리고 ${\alpha}_{S2}$-카세인, ${\alpha}_{S1}$-카세인, ${\beta}$-카세인, ${\kappa}$-카세인, ${\beta}$-락토글로불린 및 ${\alpha}$-락트알부민은 분만 직후부터 7일까지 현저한 함량의 차이는 나타나지 않았다. 한편, 돼지 초유의 경우는 모든 단백질이 분만 후, 초일부터 2일까지 높은 함량을 나타내었다.

• Reproductive and Developmental Biology
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• 제30권4호
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• pp.293-299
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• 2006

In order to generate transgenic goats expressing human growth hormone (hGH) in their mammary glands, goat ${\beta}-Casein/hGH$ hybrid gene was introduced into goat zygotes by pronuclear microinjection. DNA-injected embryos were transferred to the oviduct of recipients at 2-cell stage or to the uterus at morula/blastocyst stage after cultivation in glutathione-supplemented mSOF medium in vitro. Pregnancy and survival rate were not significantly different between 2-cell embryos and morula/blastocysts transferred to oviduct and uterus, respectively. One transgenic female goat was generated from 153 embryos survived from DNA injection. Southern blot analysis revealed that the transgenic goat harbored single-copy transgene with a partial deletion in its sequences. Despite of the partial sequence deletion, the transgene was successfully expressed hGH at the level of $72.1{\pm}15.1{\mu}g/ml$ in milk throughout lactation period, suggesting that the sequence deletion had occurred in non-essential part of the transgene for the transgene expression. Unfortunately, however, the transgene was not transmitted to her offspring during three successive breeding seasons. These results demonstrated that goat ${\beta}-casein/hGH$ gene was integrated into the transgenic goat genome in a mosaic fashion with a partial sequence deletion, which could result in a low level expression of hGH and a failure of transgene transmission.