• Asian-Australasian Journal of Animal Sciences
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• 제18권11호
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• pp.1548-1551
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• 2005

The leptin, an anti-obesity protein, is a hormone protein expressed and secreted mainly from adipocyte tissue, and involved in regulation of body weight, food intake and energy metabolism. In an effort to discover polymorphism(s) in genes whose variant(s) might be implicated in phenotypic traits of growth, we have sequenced exons and their boundaries of leptin gene including 1,000 bp upstream of promoter region with twenty-four unrelated Korean cattle. Fifty-seven sequence variants were identified: fourteen in 5' flanking region, twenty-seven in introns, eight in exons, and eight in 3' flanking region. By pair-wise linkage analysis among polymorphisms, ten sets of SNPs were in absolute linkage disequilibrium (LD) (|D'| = 1 and $r^2$ = 1). Among variants identified, thirty-six SNPs were newly identified, and twenty-one SNPs, which were reported in other breeds, were also confirmed in Korean cattle. The allele frequencies of variants were quite different among breeds. The information from SNPs of bovine leptin gene could be useful for further genetic studies of this gene.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.92-92
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• 2002

The neuropeptide vasopressin (VP) is a nine- amino acid hormone synthesized as preprohormone in the cell bodies of hypothalamic magnocellular neurons. The tumor in magnocellular neurons of the hypothalamus is associated with disfunctions of the cell bodies, leading to the diabetes insipidus. In order to produce the disease models with a defect in VP synthesis and its secretion, we have produced the transgenic mice regulated by VP constructs containing 3.8 kbp of the 5'flanking region and all the exons and introns in the mouse VP gene, which was fused at the end of exon 3 to a SV40 Tag. The two VP-transgene constructs differed by the lengths of their VP gene 3' flanking regions (2.1 versus 3.6 kbp). (omitted)

• 한국동물위생학회지
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• 제22권4호
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• pp.363-370
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• 1999

돼지 호흡기 생식기증 바이러스(porcine reproductive and respiratory syndrome virus: PRRSV) 감염이 의심되는 농장으로부터 수집된 혈청가검물 646개로부터 MARC-145 cell을 이용하여 PRRSV 분리를 시도한 바 MARC-145 세포단층상에 세포변성효과(cytopathic effects : CPE)를 나타내는 바이러스 36주를 분리하였다. 분리된 36주가 PRRSV인지 여부를 확인하기 위하여 PRRSV를 실험적으로 접종한 혈청을 이용하여 간접형광항체시험과 혈청중화시험을 실시한 결과 36주 모두가 PRRSV로 동정되었다. 혈청학적인 동정법과 더불어 reverse-transcription polymerase chain reaction을 이용하여 PRRSV open reading frame 5(ORF5)의 유전자를 증폭한 결과 선발된 6주 모두에서 80bp의 flanking sequencing를 포함하여 약 680bp의 ORF5의 유전자를 증폭할 수 있었다.

• Archives of Pharmacal Research
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• 제20권5호
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• pp.404-409
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• 1997

Trout CYP1A-CAT expression construct was generated by cloning -3.5 Kb $5^I$ flanking DNA of trout liver CYP1A gene in front of CAT gene at pCAT-basic vector. Hepa 1 cells, which are known to contain a functional arylhydrbcarbon $receptor^I$ were transfected with trout CYP1A-CAT using lipofectin. 3-Methylcholanthrene (1 nM) was added into hepa 1 cells in culture in order to examine if $5^I$ flanking DNA of trout CYP1A gene could interact with mouse transactivating factors to bring about transcription of the chloramphenicol acetyltransferase(CAT) reporter gene. The level of CAT protein was measured by CAT ELISA and the level of CAT mRNA was determined by RTPCR. The treatment of 1 nM 3-methylcholanthrene resulted in two fold increases in CAT protein as well as CAT mRNA compared to untreated control hepa 1 cells. These data indicate that arylhydrocarbon receptors of mouse hepa 1 cells are functional to activate exogenously transfected trout CYP1A-CAT construct in terms of both transcription and translation of CAT. We also examined the effect of 3-methylcholanthrene on endogenous cyplal activity in hepa 1 cell. 3-Methylcholanthrene (1 nM) treatment to hepa 1 cells trahsfected with trout CYP1A-CAT construct stimulated the level of cyp1a1 mRNA by two folds and the activity of ethoxyresorufin-O-deethylase by two fold compared to that of control cells. In this study we reported that trout CYP1A-CAT reporter gene expression construct could be expressed by 3-methylcholanthrene treatment in mouse hepa 1 cells. Thus trout CYP1A-CAT could serve as a good model to study the mechanism of regulation of CYP1A1 gene expression.

• 생명과학회지
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• 제18권4호
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• pp.461-466
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• 2008

Transforming growth $factor-{\beta}$ ($TGF-{\beta}$)에 의해 유도되는 세포사멸 과정은 정상 조직에서 손상 받은 조직이나 비정상 적인 조직을 제거하는데 중요한 역할을 담당한다. Gadd45b는 p38 kinase를 활성화시킴으로 $TGF-{\beta}$에 의해 유도되는 세포사멸 과정을 매개한다고 알려져 있다. 본 연구에서는 $TGF-{\beta}$에 의해 세포사멸이 일어나는 EpH4 세포에서 Gadd45b 유전자의 발현이 $TGF-{\beta}$에 의해 촉진됨을 보여주었다. 어떠한 기작으로 $TGF-{\beta}$에 의해 Gadd45b 유전자의 발현이 촉진되는지 알아보기 위해 Gadd45g 유전자의 5'-flanking region을 cloning하였으며, EpH4 세포에서 $TGF-{\beta}$에 의해 그 promoter activity가 증가함을 확인하였다. 여러 가지 deletion mutants를 제조하여 promoter activity를 조사한 결과 전사 개시점으로부터 220 bp upstream 부위 에 promoter activity에 필수적인 sequence가 존재함을 확인하였다. 또한 $TGF-{\beta}$에 의한 Gadd45b 유전자의 promoter activity에 Smad2, Smad3, 그리고 Smad4가 중요한 기능을 담당함도 확인하였다. 마지막으로 ras 유전자가 도입되어 $TGF-{\beta}$에 의한 세포사멸이 억제되어있는 EpRas 세포에서 $TGF-{\beta}$에 의한 Gadd45b 유전자의 발현을 확인한 결과 EpRas 세포에서 $TGF-{\beta}$에 의한 Gadd45b 유전자의 발현이 억제됨을 확인하였다. 이러한 결과는 Gadd45b 유전자가 EpH4 세포에서 $TGF-{\beta}$에 의한 세포사멸을 유도하는데 중요한 기능을 담당할 가능성이 높음을 의미하는 것이다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2000년도 국제심포지움
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• pp.1-2
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• 2000

High production of milk and its components are necessary to allow maximal growth of developing piglets. In this study, transgenic pigs were produced containing the $\alpha$-lactalbumin gene, whose product is a potential limiting component in the production of milk. Two lines of transgenic pigs were produced to analyze the effects that overproduction of the milk protein $\alpha$-lactalbumin may have on milk production and piglet growth. Transgenic pigs were produced through microinjection of the bovine $\alpha$-lactalbumin gene. The gene construct contained 2.0 kb of 5 flanking region, the 2.0 kb coding region and 329 bp of 3 flanking region. Sows hemizygous for the transgene produced as much as 0.9 g of bovine $\alpha$-lactalbumin per liter of pig milk. The production of the bovine protein caused approximately a 50 % increase in the total $\alpha$-lactalbumin concentration in pig milk throughout lactation. The concentration of bovine $\alpha$-lactalbumin was highest on day 0 and 5 of lactation and decreased as lactation progressed. The ratio of bovine to porcine $\alpha$-lactalbumin changed during the sow's lactation. This ratio was 4.3 to 1 on day 0 of lactation, but by day 20 of lactation the ratio was 0.43 to 1. This suggested that the bovine transgene and the endogenous porcine gene were under slightly different control mechanisms. The higher level of total $\alpha$-lactalbumin present on day 0 of lactation was correlated with higher lactose percentage on day 0 in transgenic sows (3.8 %) as compared to controls (2.6 %) (P < 0.01). Although there was also a trend for higher lactose percentage in transgenic sows on day 5 and 10 of lactation, no significant differences were observed. These data suggest that $\alpha$-lactalbumin is limiting early in lactation of swine. Furthermore, higher concentrations of $\alpha$-lactalbumin early in lactation may boost milk output.

• Asian-Australasian Journal of Animal Sciences
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• 제18권5호
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• pp.734-740
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• 2005

The blood samples were collected from dairy cows at the same milking stage. The single-strand conformation polymorphism (PCR-SSCP) method was used to analyze for polymorphism at the 5'flanking region of the hsp70 gene. The mRNA expression levels of HSP70, HSF1, Bcl-2 and Bax-$\alpha$ at different daily-mean-temperature were analyzed by relative quantitative RTPCR. The DNA content, cell phase and the ratio of apoptosis of lymphocytes in peripheral blood of dairy cattle at different daily-meantemperature were determined by FCM. The PCR-SSCP products of primer pair 1 showed polymorphisms and could be divided into four genotypes: aa, ab, ac, cc, with the cis-acting element (CCAAT box) included. Mutations in the hsp70 5'flanking region (468-752 bp) had different effects on mRNA expression of HSP70, HSF1, Bcl-2 and Bax-$\alpha$. The ac genotypic cows showed higher expressions of HSP70mRNA, HSF1mRNA and Bcl-2mRNA/Bax-$\alpha$mRNA and lower ratio of apoptosis. These mutation sites can be used as molecular genetic markers to assist selection for anti-heat stress cows.

• 한국가축번식학회지
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• 제24권4호
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• pp.321-333
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• 2000

High production of milk and its components are necessary to allow maximal growth of developing piglets. In this study, transgenic pigs were produced containing the $\alpha$ -lactalbumin gene, whose product is a potential limiting component in the production of milk. Two lines of transgenic pigs were produced to analyze the effects that overproduction of the milk protein $\alpha$ -lactalbumin may have on milk production and piglet growth. Transgenic pigs were produced through microinjection of the bovine $\alpha$ -lactalbumin gene. The gene construct contained 2.0 kb of 5'flanking region, the 2.0 kb coding region and 329 bp of 3'flanking region. Sows hemizygous for the trans gene produced as much as 0.9 g of bovine $\alpha$-lactalbumin per liter of pig milk. The production of the bovine protein caused approximately a 50% increase in the total $\alpha$ -lactalbumin concentration in pig milk throughout lactation. The concentration of bovine $\alpha$ -lactalbumin was highest on day 0 and 5 of lactation and decreased as lactation progressed. The ratio of bovine to porcine $\alpha$ -lactalbumin changed during the sow's lactation. This ratio was 4.3 to 1 on day 0 of lactation, but by day 20 of lactation the ratio was 0.43 to 1. This suggested that the bovine transgene and the endogenous porcine gene were under slightly different control mechanisms. The higher level of total $\alpha$-lactalbumin present on day 0 of lactation was correlated with higher lactose percentage on day 0 in transgenic sows (3.8%) as compared to controls (2.6%) (P＜0.01). Although there was also a trend for higher lactose percentage in transgenic sows on day 5 and 10 of lactation, no significant differences were observed. These data suggest that $\alpha$ -lactalbumin is limiting early in lactation of swine. Furthermore, higher concentrations of $\alpha$ -lactalbumin early in lactation may boost milk output.

• 한국응용곤충학회지
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• 제38권2호
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• pp.145-149
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• 1999

야생주 핵다각체병 바이러스의 낮은 병원성에 대해 살충제로서의 파밤나방 핵다각체병 바이러스(Spodoptera exigua nucleopolyhedrovirus: SeNPV)의 병원성 향상을 꾀함과 동시에 재조합 바이러스가 다각체 내에 매립됨으로써 살충제로의 적용시 야외에서의 안정성을 확보할 수 있는 p10 유전자의 프로모터를 이용한 새로운 발현벡터를 개발하기 위하여 국내분리주 SeNPV의 p10 유전자 구조를 분석하였다. 그 결과, SeNPV p10 유전자의 프로모터와 구조유전자 부위를 포함한 545염기서열을 결정하여 기존에 보고된 SeNPV p10 유전자(Zuidema et al., 1993)와 비교한 결과 구조유전자 부위에서는 100%의 상동성을 보였으나 5` 및 3` flanking region의 4개의 염기서열에서 차이를 보였다. Southern hybridization에 의하여 SeNPV전체 genomic DNA상에서 p10 유전자는 Sph I 2.4Kb와 Cla I 4.0Kb 단편내에 각각 존재함을 확인하였으며, p10 유전자를 포함하는 이들 단편을 각각 클로닝하여 제한효소 지도를 작성한다.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.235-235
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• 2005