• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.231-231
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• 2004

Successful embryonic development is dependant on temporal and stage-specific expression of appropriate genes. However, information on specific gene expression during early cleavage before zygotic gene activation (ZGA) is lacking. In the present study, we compared gene expression between porcine parthenotes 2-cell and blastocyst embryos to identify the genes that are specifically or prominently expressed by employing annealing control primers (ACP)-based Gene Fishing RCR. (omitted)

• Mycobiology
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• 제30권2호
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• pp.88-92
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• 2002

Rehmannia glutinosa is a perennial medicinal plant belonging to the family Scrophulariaceae with more than 300 species known in the world, especially in temperate regions. Its roots have been used widely in Korea for medicinal purposes. However, it is commonly infected by various pathogens during storage, causing great damage to the roots, and impedes the intensive farming of the crop. Therefore, an attempt has been made to isolate and screen a resistance gene against the pathogen Fusarium oxysporum using differential display. We treated salicylic acid(SA), and isolated a resistance gene that responds to SA. As a result, we found that SA was involved in plant defense mechanism in pathogenicity tests with SA treated and non-treted plants, and we isolated a partial PR-la gene through differential display polymerase chain reaction(DD-PCR) method.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권3호
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• pp.201-206
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• 2004

Vascular endothelial growth factor (VEGF), plays a key role in angiogenesis. Many endogenous factors can affect angiogenesis in endothelial cells. VEGF is known to be a strong migration, sprouting, survival, and proliferation factor for endothelial cells during angiogenesis in endothelial cells. Searching for novel genes involved in VEGF signaling during angiogenesis, we carried out differential display polymerase chain reaction on RNA from VEGF-stimulated human umbilical vein endothelial cells (HUVECs). In this study, follistatin (FS) differentially expressed in VEGF-treated HUVECs, compared with controls. Addition of VEGF (10ng/L) produced an approximately 11.8-fold increase of FS mRNA. F5 or VEGF produced approximately 1.8- or 2.9-fold increases, respectively, in matrix metalloproteinase-2 (MMP-2) secretion for 12h, compared to the addition of a control buffer. We suggest that VEGF may affect the angiogenic effect of HUVECs, through a combination of the direct effects of VEGF itself, and the indirect effects mediated via induction of FS in vitro.

• International Journal of Oral Biology
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• 제37권2호
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• pp.51-56
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• 2012

Tooth development involves bud, cap, bell and hard tissue formation stages, each of which is tightly controlled by regulatory molecules. The aim of this study was to identify genes that are differentially expressed during dental hard tissue differentiation. Sprague-Dawley rats at postnatal days 3, 6 and 9 were used in the analysis. Differential display RT-PCR (DD-PCR) was used to screen differentially expressed genes between the 2nd (root formation stage, during mineralization) and 3rd (cap stage, before mineralization) molar germs at postnatal day 9. The DNA detected in the 2nd molar germs showed homology to osteonectin only (GenBank accession no. NM_012656.1). The level of osteonectin mRNA expression was much higher in the 2nd molar germs than in the 3rd molar germs and was found to increase in a time-dependent manner from the early bell stage to the root formation stage in the 2nd molar germs. The pattern of osteonectin protein expression was consistent with these RT-PCR results. Osteonectin protein was found by immunofluorescent analysis to localize in odontoblasts and preodontoblasts rather than the dentin matrix itself. Further studies are needed to validate the involvement of osteonectin in mineralization and root formation.

• Asian-Australasian Journal of Animal Sciences
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• 제18권1호
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• pp.13-16
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• 2005

This study was performed to detect genetic variation of the heart fatty acid-binding protein (H-FABP) gene by PCRRFLPs approach and its association with intramuscular fat (IMF) content. Data from 223 individuals, including one Chinese native pig breed and four western pig breeds, were analyzed. The results showed that for the H-FABP gene, there was one polymorphic HinfI site in the 5'-upstream region, whereas there were one HaeIII and one HinfI (marked as $HinfI^*$) polymorphic site in the second intron, respectively. The three PCR-RFLPs were present in all breeds tested. The allele frequencies, however, revealed significant differences between them (p<0.05). Furthermore, the allele frequency distribution of HinfI in the Laiwu Black and that of $HinfI^*$ in the Hampshire breed were at disequilibrium, which might be the result of selective breeding. Results also indicated that for HinfI, HaeIII and $HinfI^*$ HFABP RFLP, significant (p<0.05) contrasts of 0.78%, -0.69% and 0.72% were detected in the least square means of IMF content between the homozygous genotype HH and hh, DD and dd, BB and bb classes, respectively. It implied that the HHddBB genotype had the highest IMF content in this experimental population and these H-FABP RFLPs could serve, to some extent, as genetic markers for use in improvement of IMF content.

• Journal of Ginseng Research
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• 제43권2호
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• pp.261-271
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• 2019

Background: Protopanaxatriol (PPT) is an aglycone of ginsenosides, which has high medicinal values. Production of PPT from natural ginseng plants requires artificial deglycosylation procedures of ginsenosides via enzymatic or physicochemical treatments. Metabolic engineering could be an efficient technology for production of ginsenoside sapogenin. For PPT biosynthesis in Panax ginseng, damarenediol-II synthase (PgDDS) and two cytochrome P450 enzymes (CYP716A47 and CYP716A53v2) are essentially required. Methods: Transgenic tobacco co-overexpressing P. ginseng PgDDS, CYP716A47, and CYP716A53v2 was constructed via Agrobacterium-mediated transformation. Results: Expression of the three introduced genes in transgenic tobacco lines was confirmed by Reverse transcription-polymerase chain reaction (RT-PCR). Analysis of liquid chromatography showed three new peaks, dammarenediol-II (DD), protopanaxadiol (PPD), and PPT, in leaves of transgenic tobacco. Transgenic tobacco (line 6) contained $2.8{\mu}g/g$ dry weight (DW), $7.3{\mu}g/g$ DW, and $11.6{\mu}g/g$ DW of PPT, PPD, and DD in leaves, respectively. Production of PPT was achieved via cell suspension culture and was highly affected by auxin treatment. The content of PPT in cell suspension was increased 37.25-fold compared with that of leaves of the transgenic tobacco. Transgenic tobacco was not able to set seeds because of microspore degeneration in anthers. Transmission electron microscopy analysis revealed that cells of phloem tissue situated in the center of the anther showed an abnormally condensed nuclei and degenerated mitochondria. Conclusion: We successfully achieved the production of PPT in transgenic tobacco. The possible factors deriving male sterility in transgenic tobacco are discussed.

• Asian-Australasian Journal of Animal Sciences
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• 제20권9호
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• pp.1334-1341
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• 2007

This study identified hepatic differentially expressed genes (DEGs) affecting the marbling of muscle. Most dietary nutrients bypass the liver and produce plasma lipoproteins. These plasma lipoproteins transport free fatty acids to the target tissue, adipose tissue and muscle. We examined hepatic genes differentially expressed in a differential-display reverse transcription-polymerase chain reaction (ddRT-PCR) analysis comparing high- and low-marbled Hanwoo steers. Using 60 arbitrary primers, we found 13 candidate genes that were upregulated and five candidate genes that were downregulated in the livers of high-marbled Hanwoo steers compared to low-marbled individuals. A BLAST search for the 18 DEGs revealed that 14 were well characterized, while four were not annotated. We examined four DEGs: ATP synthase F0, complement component CD, insulin-like growth factor binding protein-3 (IGFBP3) and phosphatidylethanolamine binding protein (PEBP). Of these, only two genes (complement component CD and IGFBP3) were differentially expressed at p<0.05 between the livers of high- and low-marbled individuals. The mean mRNA levels of the PEBP and ATP synthase F0 genes did not differ significantly between the livers of high- and low-marbled individuals. Moreover, these DEGs showed very high inter-individual variation in expression. These informative DEGs were assigned to the bovine chromosome in a BLAST search of MS marker subsets and the bovine genome sequence. Genes related to energy metabolism (ATP synthase F0, ketohexokinase, electron-transfer flavoprotein-ubiquinone oxidoreductase and NADH hydrogenase) were assigned to BTA 1, 11, 17, and 22, respectively. Syntaxin, IGFBP3, decorin, the bax inhibitor gene and the PEBP gene were assigned to BTA 3, 4, 5, 5, and 17, respectively. In this study, the in silico physical maps provided information on the specific location of candidate genes associated with economic traits in cattle.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9203-9210
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• 2014

Purpose: To explore associations of CYP2E1 and NAT2 polymorphisms with lung cancer susceptibility among Mongolian and Han populations in the Inner Mongolian region. Materials and Methods: CYP2E1 and NAT2 polymorphisms were detected by PCR-RFLP in 930 lung cancer patients and 1000 controls. Results: (1) Disequilibrium of the distribution of NAT2 polymorphism was found in lung cancer patients among Han and Mongolian populations (p=0.031). (2) Lung cancer risk was higher in individuals with c1, D allele of CYP2E1 RsaI/PstI, DraI polymorphisms and slow acetylation of NAT2 (c1 compared with c2, OR=1.382, 95%CI: 1.178-1.587, p=0.003; D compared with C, OR=1.241, 95%CI: 1.053-1.419, P<0.001; slow acetylation compared with rapid acetylation, OR=1.359, 95%CI:1.042-1.768, p=0.056) (3) Compared with c2/c2 and rapid acetylation, c1/c1 together with slow acetylation synergetically increased risk of lung cancer 2.83 fold. (4) Smokers with CYP2E1 c1/c1, DD, and NAT2 slow acetylation have 2.365, 1.916, 1.841 fold lung cancer risk than others with c2/c2, CC and NAT2 rapid acetylation, respectively. (5) Han smokers with NAT2 slow acetylation have 1.974 fold lung cancer risk than others with rapid acetylation. Conclusions: Disequilibrium distribution of NAT2 polymorphism was found in lung cancer patients among Han and Mongolian populations. Besides, Han smokers with NAT2 slow acetylation may have higher lung cancer risk compared with rapid acetylation couterparts. CYP2E1 c1/c1, DD and NAT2 slow acetylation, especially combined with smoking, contributes to the development of lung cancer. CYP2E1 c1/c1 or DD genotype and NAT2 slow acetylation have strong synergistic action in increasing lung cancer risk.

• 사상체질의학회지
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• 제10권2호
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• pp.283-290
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• 1998

Sasang Constitutional Medicine focuses on the different constitutional manifestations of the individual's nature and emotions. The nature and emotions drive the ascending and descending of Qi in the body. And this dynamics of the Qi's ascent and descent shapes the different types of structures, functions and temperaments. Although Sasang Constitutional Medicine has many advantages, its diagnosis of the constitution still depends on the doctors' own idea and has no objective identification. So many doctors in Korea have been trying to solve this problem. Recently, there are several efforts to find out the relationship between genetic information and constitution. By the way, May, 1998 there is a astonishing report about the gene which determines the human performance, that is ACE(angiotensin converting enzyme). And it suggests that the I allele was associated with improved endurance performance. ACE has three genotype including II, ID and DD. "I" means insertion and "D" means deletion. We determined the type of the Sasang constitution with QSCCII questionaire and the one's ACE genotype with PCR of the 127 people and we discovered the relationship between the constitution and the ACE genotype. The result is as follow. Among 39 people who have the II genotype, 7(18%) belong to Taeum(Taiyin), 9(23.1%) belong to Soyang(Shaoyang) and 23(59%) belong to Soeum(Shaoyin). Among 62 people who have the ID genotype, 18(29%) belong to Taeum(Taiyin), 21(33.9%) belong to Soyang(Shaoyang) and 23(37.1%) belong to Soeum(Shaoyin). Among 26 people who have DD genotype, 11(42.3%) belong to Taeum(Taiyin), 4(15.4%) belong to Soyang(Shaoyang) and 11(42.3%) belong to Soeum(Shaoyin). This data indicates that there are implicable relationship between the Sasang constitution and ACE genotype. Especially people who have II genotype have much possibility to be a Soeum(Shaoyin) person (59%) and Soyang(Shaoyang) person have less possibility to have DD genotype (15.4%). With this conclusion, we suggest further study of relationship between the Sasang constitution and ACE genotype and we think that other polymorphism can be a candidate of the partner of Sasang constitution.

• 대한소아치과학회지
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• 제32권1호
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• pp.75-88
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• 2005

법랑아세포종은 1868년에 처음 보고된 이래 명칭, 발생기전, 분류 그리고 치료 방법 등에 관하여 수 많은 논란이 있어 왔는데 이는 법랑세포종이 양성종양임에도 불구하고 종양자체의 진행이 파괴적이고, 외과적 처치를 한 후에도 재발이 잘되며, 흔하지는 않지만 악성종양과 유사하게 전이를 보이는 등 독특한 특성을 지니고 있기 때문이다. 정상세포와 암 세포 간에 차이를 보이는 유전자 혹은 정상세포에서 변형이 일어날 때 특이적으로 발현하는 유전의 분리 및 분석하는 것은 암세포의 생성과정을 이해하는데 있어서 중요한 열쇠를 제공할 수 있다. 이에 본 연구는 RNA differential display 방법 중 재연성과 반복성이 개선된 Ordered differential display(ODD)RT-PCR과 보다 개선된 $GeneFishing^{TM}$기술을 이용하여 악성과 양성종양 사이의 유전자 발현의 차이를 조사하고, 특이 유전자의 profile을 확보하고자 하였다. $GeneFishing^{TM}$기술과 RT-PCR을 수행한 결과 nasopharyngeal carcinoma gene을 제외한 9개의 유전자는 악성에서 특이적으로 발현되는 것을 확인하였다. 따라서 $GeneFishing^{TM}$을 이용하면 각 시료간의 mRNA 상에서 발현차이를 보이는 DEG를 비교 분석하면 암관련 유전자, 항생제 태성 유전자, 그리고 분화 관련 유전자들에 대한 연구가 용이하게 수행할 수 있을 것으로 생각된다.