• Title/Summary/Keyword: DD-PCR

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The Effect of Angiotensin Converting Enzyme Gene Polymorphism in Children with Henoch-Schonlein Purpura Nephritis (Henoch-Schonlein Purpura 신염에서 안지오텐신 전환효소 유전자 다형성의 영향)

  • Ha, Chang Woo;Kim, Ji Young;Lee, Jeong Nyeo;Lee, Jeong Hwa;Chung, Woo Yeong
    • Clinical and Experimental Pediatrics
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    • v.45 no.7
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    • pp.884-890
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    • 2002
  • Purpose : Henoch-Schonlein purpura(HSP) nephritis has been reported to vary from 25 to 50% among HSP patients and is a common cause of chronic glomerulonephritis in children. In our study, we evaluated the distribution and the association of the Insertion/Deletion(I/D) polymorphism of angiotensin converting enzyme(ACE) gene with clinical manifestations, particularly proteinuria in children with HSP nephritis, compared with that in HSP. Methods : ACE gene polymorphism was determined in children with HSP nephritis(n=33) and HSP(n=28) who were diagnosed in Busan Paik hospital from January 1996 to June 2001. The I/D polymorphism of ACE gene was determined by PCR amplication of genomic DNA. Results : The ACE I/D genotype frequency was DD : 25%, ID : 50%, II : 25% in HSP and DD : 24 %, ID : 46%, II : 30% in HSP nephritis, there was no significant difference in the genotype and allele frequencies between two groups. When statistical analysis was done according to the presence of D allele, the amount of 24-hour urinary protein excretion and the incidence of moderate to heavy proteinuria(>$500mg/m^2/day$) at onset and last follow-up were higher in DD/ID genotype than in those in II genotype, but these differences were not statistically significant. Conclusion : We suggest a lack of association between I/D polymorphism of ACE gene and clinical manifestations in children with HSP nephritis. However, further follow-up studies based on a sufficient number of patients and long term follow up periods are necessary to confirm the role of I/D polymorphism of ACE gene in children with HSP nephritis.

DD1.5k, the Gene Preferentially Expressed in Bloodstream Isolates of Vancomycin-Resistant Enterococcus faecium

  • Kim, Seung-Han;Lee, Dong-Gun;Yoo, Jin-Hong;Park, Su-Mi;Park, Jung-Hyun;Shin, Wan-Shik;Lee, Kyungwon;Dongeun Yong;Lee, Wee-Gyo
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.143-146
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    • 2004
  • Vancomycin-resistant Enterococcus faecium (VREFM) is becoming a threatening pathogen. We identified a gene called DD1.5K by differential display-PCR, which was preferentially expressed in the bloodstream isolates of VREFM. Due to its amino acid similarity to transfer complex protein, trsE, and tissue-specific expression, this gene may be involved in virulence of VREFM.

Differential display RT-PCR 기법을 이용한 돼지 등심조직의 품종 간 발현차이 유전자의 연구

  • Kim, Nam-Guk;Jo, Jung-Ho;Im, Jong-Hyeon;Bang, Gyeong-Jeong;Song, Min-Jin;Park, Beom-Yeong;Kim, Eon-Hyeon;Lee, Chang-Su
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2005.05a
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    • pp.239-242
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    • 2005
  • 본 연구는 성장 속도 및 서로 다른 육질 특성을 지닌 돼지 품종을 이용하여, 육질 및 성장에 관련된 유전자원을 확보하고, 이를 이용한 유전 육종의 기초 자료를 제공하기 위하여 수행하였다. Differential display (DD) RT-PCR 기법을 통해 돼지 품종 간 발현 차이를 보이는 유전자인 NADH dehydrogenase 1과 ATPase 6를 동정하였다. 동정된 유전자의 발현량 분석을 위한 RT-PCR 결과, 각 유전자의 발현량이 재래돼지에서 외래 품종 (랜드레이 스 및 요크셔)에 비해 2배 이상 높음을 확인 할 수 있었다 (p<0.01). 이러한 발현차이 유전자를 이용하여 육질과의 관련성 연구 및 유전자의 기능에 대한 연구가 지속되어야 할 것이다.

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Age-dependent Changes of Differential Gene Expression Profile in Backfat Tissue between Hybrids and Parents in Pigs

  • Ren, ZH.Q.;Xiong, Yuanzhu;Deng, CH.Y.;Zuo, B.;Liu, Y.G.;Lei, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.682-685
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    • 2005
  • Large White, an introduced European pig breed, and Meishan, a Chinese indigenous pig breed, were hybridized directly and reciprocally and a total of 260 pigs, including purebreds, Large White and Meishan, and their hybrids, White${\times}$Meishan (LM) and Meishan${\times}$Large White (ML) pigs, were bred in our laboratory. The mRNA differential display PCR (DD-PCR) was used to detect the age-dependent changes of differential gene expression in backfat tissue between hybrids and parents. Some measures were taken to reduce the false positives in our experiment. Among the total of 2,686 bands obtained, 1,952 bands (about 72.67%) were reproducible and eight patterns (fifteen kinds) of gene expression were observed. The percentage of differentially expressed genes between hybrids and parents is 56.86% at the age of four months and 57.71% at the age of six months. This indicated that the differences of gene expression between hybrids and their parents were very obvious. U-test was used to compare the patterns of gene expression between the age of four and six months, and results showed that bands occurring in only one hybrid and bands displayed in one hybrid and one parent were significantly different at p<0.05, and bands visualized in only two hybrids were significantly different at p<0.01. These indicated that differential gene expression between hybrids and parents changed at different ages.

Differential Expression of Ubiquitin-Specific Protease 16 Gene by Methylprednisolone in Neuronal Cells

  • Choi, Seung-Won;Kwon, Hyon-Jo;Koh, Hyeon-Song;Song, Shi-Hun;Kwon, O-Yu;Kim, Seon-Hwan
    • Biomedical Science Letters
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    • v.16 no.2
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    • pp.105-112
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    • 2010
  • Methylprednisolone (MPD) is a synthetic glucocorticoid drug used in treatment of many neurological diseases and neurotraumas, including spinal cord injuries. Little is known of the mechanism of MPD in neuronal cells, particularly the genetic expression aspect. DD-PCR was used in identification of genes expressed during MPD treatment of PC12 cells. We have isolated 3 predicted up- or down-regulated genes, which are differentially expressed in neurons by MPD. One of these genes, USP16 (ubiquitin specific protease 16), is the deubiquitinating enzyme that is up-regulated by MPD in neurons. In order to observe the effect of MPD on USP16 gene expression, PC12 cells were treated under several experimental conditions, including endoplasmic reticulum stress drugs. We have isolated the total RNAs in PC12 cells and detected USP16 and ER related genes by RT-PCR. Because its expression pattern is similar to expression of ER chaperons, USP16 gene expression is strongly associated with unfolded protein response. A meaningful negative effect on each tissue treated by methylprednisolone is not shown in vivo. USP16 gene expression is suppressed by LY294002 (phosphatidylinositol 3-kinase inhibitor), which suggests that USP16 gene expression is regulated by the phosphatidylinositol 3-kinase pathway.

Isolation and Characterization of Defense Genes Mediated by a Pathogen-Responsive MAPK Cascade in Tobacco (담배에서 병원균에 반응하는 MAPK 신호전달체계에 의해 매개되는 방어 유전자들의 분리 및 특성화)

  • Jang, Eun-Kyoung;Kang, Eun-Young;Kim, Young-Cheol;Cho, Baik-Ho;Yang, Kwang-Yeol
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1023-1030
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    • 2008
  • NtMEK2, which is the tobacco MAPK kinase that is upstream of SIPK and WIPK, was identified using the dexamethasone (DEX)-inducible gain-of-function transgenic system. Expression of $NtNEK2^{DD}$, a constitutively active mutant of NtNEK2, leads to HR-like cell death, which indicates that the NtMEK2-SIPK/WIPK cascade controls defense responses in tobacco. However, little is known about the downstream target substrates or defense-related genes that are regulated by the NtMEK2-SIPK/ WIPK cascade. In this study, ACP-based differential display RT-PCR was used to isolate the downstream effectors mediated by the NtMEK2-SIPK/WIPK cascade in $NtNEK2^{DD}$ transgenic plants. The results identified 6 novel differentially expressed genes (DEGs). These included pathogen induced protein 2-4 (pI2-4), monoterpene synthase 2 (MTS2), seven in absentia protein (SINA), cell death marker protein 1 (CDM1), hydroxyproline-rich glycoprotein (HRGP) and unknown genes (DEG45). The induction of these genes was confirmed by RT-PCR of samples obtained from $NtNEK2^{DD}$ plants. Additionally, when compared with other isolated DEGs, the pI2-4, CDM1 and HRGP genes were significantly up-regulated in response to treatment with salicylic acid and tobacco mosaic virus. Taken together, these results suggest that three novel DEGs were regulated by the NtMEK2-SIPK/WIPK cascade involved in disease resistance in tobacco.

Analysis of Carcass Characteristics in the 3rd Intron of Pig POU1F1 Gene (돼지의 POU1F1 Intron 3영역 유전자에 따른 도체특성 분석)

  • Kim, Gye-Woong;Yoo, Jae-Young
    • Journal of Animal Science and Technology
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    • v.51 no.4
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    • pp.283-288
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    • 2009
  • This study was carried out to compare Msp I polymorphisms in the 3rd intron of porcine gene encoding the pituitary-1 transcription factor (POU1F1) from 286 pigs (Landrace $\times$ Yorkshire $\times$ Duroc, LYD) and to determine the associations between its genotypes and carcass traits by using the PCR-RFLP technique. The frequency of the single nucleotide polymorphism (SNP) genotype DD (84.33%) was very higher than that of CC genotype (0.75%). Allelic frequencies for C and D were 0.082 and 0.918, respectively. Each population followed the Hardy-Weinberg equilibrium. Meat colours of Hunter $L^*$ values and visual colour according to two genotypes were all significantly different. However, no significant difference in crossbred (LYD) was found between CD and DD genotypes for other traits. Therefore, this suggests that POU1F1 may be a major gene or marker for carcass traits.

Differentially expressed genes in sex determination of zebrafish

  • Lee, Jae-Hyun
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 1998.07a
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    • pp.61-62
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    • 1998
  • The effects of rearing temperature on the sex determination of zebrafish were examined to address the questions, "Is it possible to control phenotypic sex artificially; and what genes may be involved in that process\ulcorner". At higher temperature(${32-34}^{\circ}C vs 28.5^{\circ}$C), a dramatic shift of the sex ratio(female 1 : male 9) was found. with DD-PCR cloning, several candidate genes were cloned and analyzed. Further analysis should help to understand sex-determining mechanisms involved in the artificial induction.induction.

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