• 한국축산식품학회지
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• 제18권2호
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• pp.185-191
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• 1998

Human milk and bovine milk in normal stage were fractionated four parts : whey, skimmilk membrane, and casein pellet. The specific activity (nmole / mim / mg protein) and distribution ratio(%) of suborganella marker enzymes in each separated milk fraction were determined. Especially, neutral $Ca^{2+}$-ATPase, acid $Ca^{2+}$-ATPase, NADH-cytochrome C reductase, and acid phosphatase were higher in human milk. However, both $Ca^{2+}$-ATPases were not detected in all fractions of bovine milk. On the other hand, 5'-nucleotidase, phosphodiesterase I, alkaline phosphatase, and $\gamma$-glutamyl transpeptidase activities in bovine milk were higher than in human milk. Most of the marker enzymes were highly distributed in cream fraction of either human milk or bovine milk, and their specific activities were high to 24 fold from 3 fold when compared with that of whole milk. These results suggest that marker enzymes in mammary epitherial cell are transfered into cream fraction by the membrane rearrangement, and different biochemical reaction between human and bovine exists for milk secretion in mammary gland.

• KSBB Journal
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• 제17권1호
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• pp.106-109
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• 2002

Gel electrophoresis of DNA is a well known technique in molecular biology. This technique is simple, rapid to perform, and capable of adequately separating fragments of DNA. A number of mixtures of DNA fragments ("DNA size markers") are frequently employed in a purpose of extrapolating the sizes or the amount of DNA molecules during gel electrophoresis. DNA size markers are constructed by digesting plasmid DNA, bacteriophage DNA, or recombinant DNA molecules with one or more restriction enzymes. However, liquid suspension containing DNA size marker needs to be kept at a low temperature during storage and shipping. In an attempt to maintain the DNA samples at room temperature for extended period of time, lyophilization of DNA with addition of nuclease inhibitor was studied. Gel loading buffer was also added to the lyophilized DNA to provide additional convenience such that DNA size marker was the "ready-to-use" followed by simply reconstituting with distilled water.

• 한국축산식품학회지
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• 제25권2호
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• pp.218-225
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• 2005

본 연구는 mt DNA의 D-loop 영역을 probe로 이용한 Southern blot hybridization 분석 기법을 이용하여 한우에서 mt DNA의 RFLP를 분석하고 RFLP marker가 유생산에 미치는 영향을 분석하여 산유량 관련 DNA marker를 개발하고자 수행하였다. Mt DNA의 D-loop 영역내 404번부터 15061까지 1142 bp 크기의 염기 서열 부위를 특이적인 primer를 이용하여 PCR로 증폭하였다. Mt DNA를 HpaII, BamHI, XbaI, HinfI, EcoRI, HindII 및 RsaI 7종류의 제한효소를 이용하여 각각 절단한 후 DIG로 표지된 D-loop probe를 이용하여 검출한 결과 XbaI, RsaI, BamHI 및 HpaII 4종류의 제한효소에서 각각 RFLP 다형성이 검출되었고 EcoRI, HindIII 및 HinfI 3종류 제한효소는 변이가 존재하지 않았다. 다유 계통과 저유 계통 선발 집단간의 각 제한효소별 RFLP type의 출현빈도를 비교한 결과 BamHI 및 RsaI 제한효소에서 두 집단간의 RFLP type의 출현율에 각각 통계적 유의성(P<.05)이 인정되었다. 다유 및 저유성으로 극단의 육종가 값을 갖는 두 계통의 mt DNA D-loop영역의 염기 서열을 비교 분석한 결과 441번째 염기가 G/C, 469번째 염기는 T/C, 503번째 염기는 C/T, 569번째 염기는 G/A, 614번째 염기는 C/A그리고 644번째 염기는 C/T로 각각 염기가 치환되었고 특히, 다유 계통 개체의 677번째의 A염기가 저유 계통 개체에서는 결실되어 있다는 사실이 확인되었다. 한편, 한우 암소의 유생산에 영향을 미치는 효과를 규명하기 위하여 mt DNA RFLP 형과 송아지 이유시 체중, 생시체중 및 비유량을 측정하여 얻어진 육종가의 성적을 근거로 통계 분석한 결과 XbaI 제한효소의 RFLP type이 산유 능력 육종가와 유의적인 관련성이 확인되었다 (P<05). 즉, RFLP A type을 갖는 축군의 평균 육종가 추정치가 6.233으로 B type을 갖는 축군의 평균 육종가 추정치 0.757보다 월등히 높은 것으로 나타났다. 결론적으로 산유량과 관련성이 확인된 mt DNA RFLP type은 한우의 산유량 향상을 위한 DNA marker로 이용 가능할 것으로 기대된다.

• 생명과학회지
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• 제28권11호
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• pp.1277-1284
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• 2018

본 연구에서는 효모염색체내에 다양한 유전자 발현 cassette를 도입하기 위해 Cre/loxP system을 가진 repeated yeast integrative plasmid (R-YIp)를 구축하였다. R-YIp는 반복적으로 형질전환체를 선별할 수 있는 selective marker (CgTRP1)와 loxP 서열, 그리고 integration을 위한 목적서열을 함유하고 있어 같은 염색체의 동일한 위치에 여러 개의 유전자 발현 cassette를 도입하는 것이 가능하다. 따라서 xylan/xylose 대사에 관련된 endoxylanase (XYLP), ${\beta}$-xylosidase (XYLB), xylose reductase (GRE3) 그리고xylitol dehydrogenase (XYL2)의 효모염색체내에 도입을 시도하였다. 먼저 XYLP, XYLB, GRE3그리고 XYL2 유전자의 효율적인 발현을 위한 promoter를 선별하기 위해 pGMF-GENE과 pAMF-GENE plasmid를 구축하였고, 각 유전자들의 발현에 GAL10 promoter가 적합함을 확인하였다. 다음으로 GAL10p-GENE-GAL7t cassette를 가진 pRS-GENE plasmid (R-YIp)를 구축하여, 반복적 integration 과정과 selective marker의 제거를 통해 각각의 R-YIps를 효모 7번염색체에 순차적으로 도입하였다. R-YIp system을 통해 효모염색체내에 도입된 유전자들은 모두 안정적으로 발현되었고, 활성형의 재조합효소를 생산함을 확인할 수 있었다. 따라서 다수의 외래유전자를 효모염색체내 도입함에 있어 selective marker와 숙주세포 선택의 한계를 R-YIp system을 통해 어느 정도 극복할 수 있을 것이라 기대한다.

• BMB Reports
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• 제42권10호
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• pp.661-666
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• 2009

Porcine pancreas development is not well studied at the molecular level despite being a therapeutic resource for diabetic patients. In this study, we investigated expression of lineage markers and performed proteomic analysis. Expression of the early lineage markers Pdx1 and Ptf1a was developmentally conserved between mice and pigs, whereas expression of the islet differentiation marker Pax4 was delayed in porcine compared with murine pancreas development. Proteomic analysis found that expression levels of chymotrypsinogen were down-regulated during porcine pancreas development while those of digestive enzymes like lipases, elastase and serine protease were up-regulated. In addition, specific isoforms of protein folding assistants such as protein disulfide isomerase and prefoldin were expressed at specific stages during the maturation of digestive enzymes. Taken together, these results show that development of the porcine pancreas is regulated by a concerted interplay of pancreas lineage marker proteins and other specified proteins, resulting in a functional endocrine and exocrine organ.

• Preventive Nutrition and Food Science
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• 제8권4호
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• pp.365-371
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• 2003

The potential of seven flavonoid glycosides to induce quinone reductase (QR), an anticarcinogenic marker enzyme, in murine hepatoma cells (hepalc1c7) and its mutant cells (BPRc1) was evaluated. Among test compounds, kaempferol-3-O-glucoside, luteolin-6-c-glucoside, and quercetin-3-O-glucoside (Q-3-G) induced QR in hepalc1c7 cells in a dose-dependent manner. However, in BPRc1 cells lacking arylhydrocarbon receptor nuclear translocator (ARNT), only Q-3-G caused a significant induction of quinone reductase at the concentration range of 0.5 to 8 ug/mL, suggesting that it is a monofunctional inducer. Q-3-G induced not only phase 2 enzymes, including QR and glutathione-S-transferase, but also nitroblue tetrazolium reduction activity in HL-60 cells, a biochemical marker for cell differentiation promoting agents. In conclusion, Q-3-G merits further study to evaluate its cancer chemopreventive potential.

• 한국작물학회지
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• 제57권1호
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• pp.71-77
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• 2012

The objective of this study is to develop the gene based sequence tagged site (STS) markers in wheat. The euchromatin enriched genomic library was constructed and the STS primer sets were designed using gene based DNA sequence. The euchromatin enriched genomic (EEG) DNA library in wheat was constructed using the $Mcr$A and $Mcr$BC system in $DH5{\alpha}$ cell. The 2,166 EEG colonies have been constructed by methylated DNA exclusion. Among the colonies, 606 colonies with the size between 400 and 1200 bp of PCR products were selected for sequencing. In order to develop the gene based STS primers, blast analysis comparing between wheat genetic information and rice genome sequence was employed. The 227 STS primers mainly matched on $Triticum$ $aestivum$ (hexaploid), $Triticum$ $turgidum$ (tetraploid), $Aegilops$ (diploid), and other plants. The polymorphisms were detected in PCR products after digestion with restriction enzymes. The eight STS markers that showed 32 polymorphisms in twelve wheat genotypes were developed using 227 STS primers. The STS primers analysis will be useful for generation of informative molecular markers in wheat. Development of gene based STS marker is to identify the genetic function through cloning of target gene and find the new allele of target trait.

• Biomolecules & Therapeutics
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• 제31권6호
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• pp.619-628
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• 2023

In the modern era, chronic kidney failure due to diabetes has spread across the globe. Prunetin (PRU), a component of herbal medicines, has a broad variety of pharmacological activities; these may help to slow the onset of diabetic kidney disease. The anti-nephropathic effects of PRU have not yet been reported. The present study explored the potential nephroprotective actions of PRU in diabetic rats. For 28 days, nephropathic rats were given oral doses of PRU (20, 40, and 80 mg/kg). Body weight, blood urea, creatinine, total protein, lipid profile, liver marker enzymes, carbohydrate metabolic enzymes, C-reactive protein, antioxidants, lipid peroxidative indicators, and the expression of insulin receptor substrate 1 (IRS-1) and glucose transporter 2 (GLUT-2) mRNA genes were all examined. Histological examinations of the kidneys, liver, and pancreas were also performed. The oral treatment of PRU drastically lowered the blood glucose, HbA1c, blood urea, creatinine, serum glutamic-oxaloacetic transaminase, serum glutamic pyruvic transaminase, alkaline phosphatase, lipid profile, and hexokinase. Meanwhile, the levels of fructose 1,6-bisphosphatase, glucose-6-phosphatase, and phosphoenol pyruvate carboxykinase were all elevated, but glucose-6-phosphate dehydrogenase dropped significantly. Inflammatory marker antioxidants and lipid peroxidative markers were also less persistent due to this administration. PRU upregulated the IRS-1 and GLUT-2 gene expression in the nephropathic group. The possible renoprotective properties of PRU were validated by histopathology of the liver, kidney, and pancreatic tissues. It is therefore proposed that PRU (80 mg/kg) has considerable renoprotective benefits in diabetic nephropathy in rats.

• Korean Journal of Acupuncture
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• 제18권1호
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• pp.21-26
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• 2001

발암물질을 무독화시키는 QR 생성 유도를 살펴보기 위하여 황기 약침액 및 열수추출액을 생쥐의 간암세포인 Hepa1c1c7에 처리하여 측정한 결과, 황기 약침액의 농도를 증가시킬수록 많은 QR 생성율을 보였으며, GSH 생성이 증가하였고, GST 생성 또한 증가하였다.

• 약학회지
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• 제38권1호
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• pp.12-19
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• 1994

Capsaicin(8-methyl-N-vanillyl-6-nonenamide) is the principal pungent component of Capsicum fruits. This work is directed to the capsaicin-hydrolyzing enzyme playing a key role in the rate limiting and critical step of capsaicin metabolism. In order to get precise information on the enzyme's subcellular location, rat liver homogenate was divided into six subcellular fractions by differential centrifugation technique: crude nuclear pellet, PNS(post nuclear supernatant) fraction, lysosomal pellet, cytosol, Tris wash fraction, micrisomes. Capsaicin-hydrolysing enzyme activity was analysed by high performance liquid chromatography(HPLC). This enzyme was found at the highest specific activity in the microsomal fraction and co-distributed with marker enzymes of the endoplasmic reticulum, NADPH-cytochrome c reductase and nucleoside diphosphatase. This is compatible with the result of ninhydrin color reaction of vanillylamine, primary metabolite of capsaicin hydrolysis, on thin layer chromatography(TLC). This enzyme is most active at pH $8.0{\sim}9.0$. Definite subcellular location of this enzyme will make it easy to proceed with further study.