• 제목/요약/키워드: RT-PCR (reverse transcription-polymerase chain reaction)

검색결과 644건 처리시간 0.033초

A STUDY ON THE EXPRESSION OF TYPE I AND TYPE II COLLAGEN GENES AND PROTEINS IN THE DEVELOPING HUMAN MANDIBLE

  • Kook, Yoon-Ah;Kim, Sang-Cheol;Kim, Eun-Cheol
    • 대한치과교정학회지
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    • 제25권6호
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    • pp.723-731
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    • 1995
  • Type I and type II collagens are considered the major collagens of bone and cartilage respectively. Monitoring the patterns of those gene and protein expressions during development will provide a basis for the understanding of the normal and abnormal growths. This study was undertaken to investigate the expression of collagen genes and proteins involved in the developing human mandible. Fifty embryos and fetuses were studied with Alcian blue-PAS, Masson's Trichrome, reverse transcription polymerase chain reaction (RT-PCR), Western blot analysis, and Southern blot analysis. Our results showed that $pro-{\alpha}1(II)$ collagen gene expression begins in the 5th week. Type II collagen is synthesized in mesenchymal cells in advance: of overt chondrogenesis. The gene expression for type II collagen was highest during the appearance of Meckel's cartilage. There was a switch in collagen protein expression from type I to type II during the appearance stage of Meckel's cartilage. The distribution of the mRNA for type II collagen corresponded well with the pattern of type II collagen protein. The endochondral ossification was observed where there was direct replacement of cartilage by bone.

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효모시스템에서 Human Transglutaminase C(TGase II)의 발현에 관한 연구 : C-말단부위의 결손효과

  • 우상규;정선미;이상기;안병윤;김희철
    • 한국미생물·생명공학회지
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    • 제24권3호
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    • pp.290-298
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    • 1996
  • In an effort to understand the role of the conserved domain and of the heterologous one-third part of the carboxy terminal domain of transglutaminase C (TGase II), attempts were made to express TGase II cDNA of human origin in yeast Saccharomyces cerevisiae as in a full-length form as well as in a form of C-terminal truncation. The 2$\mu$-based expression plasmids which contained the TGase II cDNA under the gal inducible promoter were introduced into yeast and the maintenance of the full-length and truncated form of the TGase II gene plasmids were confirmed by Southern blot. The expression of the TGase II gene was analysed by reverse transcription polymerase chain reaction (RT-PCR), and western blot analyses. As assayed by [1,4$^{14}$C]-putrescine incorporation into succinylated casein, the full-lenth as well as the truncated forms of recombinant TGase II showed some catalytic activity. These results indicate that the N-terminal homologous domain of human TGase II retains a catalytically active domain. The level of TGase II expressed in yeast, however, was far lower than satisfactory and other expression system should be sought further chracterization of the enzyme. The negative effect of TGase II on the growth of yeast is interesting with respect to the physiological effect of TGase II in cornification of epidermal keratinocytes.

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Selective Gene Express Profiles in Rat Uterus during Estrus Cycle

  • Kim, Do-Rim;Yu, Seong-Jin;Kim, Jee-Yun;Youm, Mi-Young;Lee, Chae-Kwan;Kang, Sung-Goo
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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    • pp.70-70
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    • 2003
  • The uterus undergoes dynamic changes during the cycle and displays many features typical of developmental process. In order to be prepared for implantation, endometrium undergoes predictable, sequential phases of proliferation and secretory changes. The uterus during estrus cycle synthesize a complex of signaling molecules with specific spatial and temporal modes of expression and which are critical for cell proliferation and differentiation. The purpose of this investigation was to use cDNA microarrays to evaluate the expression of genes of rat uterus in estrus cycle. Animals were sacrificed on proestrus, estrus, metestrus, diestrus. Differential gene expression profiles were revealed(growth-related c-myc reponsive protein RCL, heat shock 47-kDa protein (HSP47), cytochrome c oxidase polypeptide Vlc2 (COX6C2), calreticulin (CALR)). Reverse transcription polymerase chain reaction (RT-PCR) was used to validate the relative expression pattern. Using this approach, we found several genes whose expression in rat uterus was altered with estrus cycle. Our long-term goal is to determine the role of these differentially expressed genes during estrus cycle. This study was supported by through the Biohealth Products Research Center(BPRC), Inje University.

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Molecular Parameters for Assessing Marine Biotoxicity: Gene Expressions of Rockfish (Sebastes schlegeli) Exposed to Polycyclic Aromatic Hydrocarbons

  • Woo, Seon-Ock;Yum, Seung-Shic;Park, Hong-Seog;Jung, Jee-Hyun;Lee, Suk-Chan;Kim, So-Jung;Lee, Taek-Kyun
    • Molecular & Cellular Toxicology
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    • 제3권4호
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    • pp.267-272
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    • 2007
  • Environmental and anthropogenic changes affect the health and stability of marine ecosystem. In this study we aimed to identify molecular biomarkers for ecotoxicological pollutants risk assessment in the rockfish (Sebastes schlegeli). We designed primers based on conserved sequences by multiple alignments of target genes from related species, and cloned the partial cDNAs of cytochrome P450 (CYP1A1), glutathione S-transferase (GST), metallothionein (MT), superoxide dismutase (SOD), ubiquitin (UB), vitellogenin (VTG) and $\beta$-actin by reverse transcription polymerase chain reaction (RT-PCR) from S. schlegeli. Northern blot results indicated that these six genes expressions were significantly induced by benzo[a]pyrene (BaP, 1 ${\mu}M$) and that the level of each of their transcripts increased in BaP-exposed rockfish in a time-dependent manner. This study suggests that transcriptional changes in these six genes may be used for monitoring environmental exposure to polycyclic aromatic hydrocarbons (PAHs).

Heme Oxygenase-1(HO-1) induction by UVB(290-320nm) radiation in ICR mice

  • Choi, Wook-Hee;Kim, Tae-Hwan;Ahn, Ryoung-Me
    • 한국환경보건학회:학술대회논문집
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    • 한국환경보건학회 2005년도 가을학술대회
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    • pp.166-168
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    • 2005
  • The induction of heme oxygenase-1(HO-1) by ultraviolet(UV) radiation provides a protective defense against oxidative stress, and has been well demonstrated in UVA-irradiated skin, but not UVB. In this study in mice, we show that the UVB(290-320nm) radiation can be attributed to the induction of cutaneous heme oxygenase-1. The expression of HO-1 mRNA was assessed in vivo by the reverse transcription-polymerase chain reaction (RT-PCR) analysis, and HO-1 enzyme activity was measured in microsomal preparation from irradiated mice. The mRNA level of HO-1 increases in liver and skin from 24h to 72h after UVB($3KJ/m^3$) radiation. The results of gene expression were same pattern of HO enzyme activity in skin, but not in liver. HO-1 mRNA in liver resulted in a progressive increase to 96h after UVB radiation, but HO activity in liver increased to 48h. This finding indicates that UVB radiation is an important inducer of HO-1 and increases in HO activity may protect tissues directly or indirectly from oxidative stress.

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ICR mouse의 피부와 간에서 자외선 B파에 의한 Heme Oxygenase-1의 유도 (Heme Oxygenase-1 (HO-1) Induction by UVB (290-320nm) Radiation in ICR Mice)

  • 최욱희;김태환;안령미
    • 한국환경보건학회지
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    • 제34권1호
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    • pp.49-54
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    • 2008
  • The induction of heme oxygenase-1(HO-1) by UV radiation provides a protective defence against oxidative stress, and has been well demonstrated in skin irradiated with UVA, but not UVB. In this study, we show that the induction of cutaneous HO-l can be attributed to UVB radiation. The expression of HO-1 mRNA was assessed in vivo by reverse transcription-polymerase chain reaction (RT-PCR) analysis, and HO-1 enzyme activity was measured in microsomal preparation from irradiated mice. The mRNA level of HO-1 increases in liver and skin from 1d to 3d after UVB $(3KJ/m^2)$ exposure. The results of gene expression were same pattern of HO-1 enzyme activity in skin, but not in liver. HO-1 mRNA in liver resulted in a progressive increase to 4d after UVB exposure, but HO-1 activity in liver increased to 2d. This finding indicates that UVB radiation is an important inducer of HO-1 and increases in HO activity may protect tissue directly or indirectly from oxidative stress.

Differentially Expressed Genes in Porcine Parthenogenetic 2-cell and Blastocyst Embryos

  • Lee, Hwa-young;Kim, Bong-Ki;Cui, Xiang-Chun;Hwang, Kyu-Chan;Chung, Hak-Jae;Ko, Yeoung-Gyu;Jung, Jin-Kwan;Chang, Won-Kyong;Kim, N. H.
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2004년도 춘계학술발표대회
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    • pp.198-198
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    • 2004
  • The identification of embryo-specific genes would provide insights into early embryonic development. However, the current methods employed to identify the genes that are expressed at a specific developmental stage are labor intensive and suffer from high rates of false positives. Here we employed a new and accurate reverse transcription-polymerase chain reaction (RT-PCR) technology that involves annealing control primers(ACPs) to identify the genes that are specifically expressed in porcine blastocysts compared to 2-cell stage embryos. (omitted)

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Photoprotective Effect of Topical EGb 761 and Korean Red Ginseng in C57BL/6 Mice

  • Choi, Wook-Hee;Han, Seon-Kyu;Yi, Seh-Yoon;Ann, Hyoung-Soo;Ahn, Ryoung-Me
    • 한국환경보건학회:학술대회논문집
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    • 한국환경보건학회 2005년도 국제학술대회
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    • pp.306-310
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    • 2005
  • Exposure to UVB radiation can cause diverse biological photodamage to skin. Eeb 761 and Korean red ginseng are the major and most effective natural drug against a variety of oxidative damage. But, the protective effects against UVB radiation have not been clearly identified. In this study, we investigated the protective effect of topical EGb 761 and Korean red ginseng on pigmentation by UVB radiation. Pro-inflammatory cytokines($IL-l{\beta}$, IL-6 and $TNF-{\alpha}$) and melanogenesis proteins(tyrosinase, TRP-1 and TRP-2) mRNA were measured by reverse transcription-polymerase chain reaction(RT-PCR) analysis. The in vivo protection against pigmentation was calculated using chromameter. The mRNA level of IL-lf and TNF-a were increased by UVB irradiation in treated and non-treated group, while no significant changes were observed in IL-6 level. Topical treatment with EGb 761 and Korean red ginseng remarkably reduced expression of tyrosinase, TRP-1 and TRP-2 in the non-irradiated and irradiated skin. Application of EGb 761 and Korean red ginseng significantly protected the WB-induced skin pigmentation and Korean red ginseng was more effective. Our study suggests that topical ECb761 and Korean red ginseng can regulate melanogenic proteins and protect UVB radiation on skin pigmentation.

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계지복령환이 Cholesterol 식이 생쥐의 SOD 유전자발현능에 미치는 영향 (Effects of geagibokrounghwan on superoxide dismutase gene expression in experimental mouse fed cholesterol)

  • 홍윤표;노승현
    • 대한한의학방제학회지
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    • 제6권1호
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    • pp.215-226
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    • 1998
  • Geagibokrounghwan (桂技茯笭湯) has long been used to cure human diseases such as vascular and blood disorders. However, it is still unkown on its action mechanism, physiolosical and biochemical meaning. Thus, many attempts were tried to show the scientific background covering the above mentioned mechanism. The effect of Geagibokrounghwan, which was known to date, as follow; effective circulation of body blood system, proliferation of leucocytes and antioxidative action. In this study, we have applied the Geagibokrounghwan administration and feed to mouse, to see effects on the expression of superoxide dismutase(SOD) mRNA as antioxidative agent and oxygen radical scavenger. Total RNAs includingmRNA have been isolated from liver and white blood cells after mice were fed with cholesterol in high dose. Also, in a separate group, the cholesterol-administrated mice were fed with Geagibokrounghwan to see the effects on SOD transcription. and then reverse transcriptase-polymerase chain reaction (RT-PCR) usion each primer set (SOD-F;GATGAAAGCGGTGT-3'; SOD-R; 5'-CCTGTGGAGTGATT-3') were performed to trace theamounts of mRNA. SOD mRNA was specifically expressed in Geagibokrounghwan-fed mice at 2 weeks after treatment, however, gradually reduced after 4 weeks. These results indicate that Geagibokrounghwan is highly applicable in treatment of the above mentioned human diseases.

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The Protective Effect of Paeoniae Radix Extract against 1-methyl-4-Phenylpyridium-induced Apoptosis on SK-N-MC Neuroblastoma Cells

  • Chae, Ki-Heon;Song, Yun-Kyung;Lim, Hyung-Ho
    • 대한한의학회지
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    • 제26권4호
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    • pp.74-86
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    • 2005
  • Background: There are increasing neuro-degenerative disorders with aging. Paeoniae Radix(PR) possesses various pharmacological effects such as sedative, analgesic, anti-inflammatory, anti-stress and neuro-protective actions. Also antiaging and anti-cancer effects of PR were reported. Our purpose was to investigate whether PR is useful on the treatment of Parkinson's disease, one of the neuro-degenerative disorders. Objective: We investigated whether Paeonia Radix possesses a protective effect against 1-methyl-4 phenylpyridine(MPP+)-induced cytotoxicity in neuronal cells. Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, flow cytometry, DNA fragmentation assay, reverse transcription-polymerase chain reaction(RT-PCR), and Western blotting were performed on SK-N-MC neuroblastoma cells. Results: Cells treated with MPP+ exhibited several apoptotic features, while cells pre-treated with Paeonia Radix prior to MPP+ exposure showed s decrease in the occurrence of apoptotic features. Conclusions: These results suggest that Paeonia Radix may exert a protective effect against MPP+-induced apoptosis in SK-N-MC neuroblastoma cells.

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