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조화자,Jeong Min Shin,김종수,Man Ryul Lee,홍기성,Jun-Ho Lee,Kyoung Hwa Koo,박정우,김계성 한국분자세포생물학회 2009 Molecules and cells Vol.28 No.6
Previously, we have reported tissue- and stage-specific expression of miR-372 in human embryonic stem cells and so far, not many reports speculate the function of this microRNA (miRNA). In this study, we screened various human cancer cell lines including gastric cancer cell lines and found first time that miR-372 is expressed only in AGS human gastric adenocarcinoma cell line. Inhibition of miR- 372 using antisense miR-372 oligonucleotide (AS-miR-372) suppressed proliferation, arrested the cell cycle at G2/M phase, and increased apoptosis of AGS cells. Furthermore, AS-miR-372 treatment increased expression of LATS2, while over-expression of miR-372 decreased luciferase reporter activity driven by the 3′ untranslated region (3′ UTR) of LATS2 mRNA. Over-expression of LATS2 induced changes in AGS cells similar to those in AGS cells treated with AS-miR-372. Taken together, these findings demonstrate an oncogenic role for miR-372 in controlling cell growth, cell cycle, and apoptosis through down-regulation of a tumor suppressor gene, LATS2.
한국에서 분리된 전염성 조혈괴저 바이러스의 non-virion (NV) 단백질의 유전자 클로닝 및 바이러스 증식에서의 역할
문창훈,조화자,윤원준,박정재,박정민,김현주,도정완,이주양,임채렬 한국미생물학회 2000 미생물학회지 Vol.36 No.2
한국에서 분리된 전염성 조혈괴저바이러스(infectious hematopoietic necrosis virus, IHNV)인 IHNV-PRT의 non-viron(NV)단백질을 암호화하고 있는 cDNA를 클로닝하여 이들의 염기서열을 분석하였다. NV는 336bpzmrl의 open reading frame을 포함하였으며 이로부터 111개의 아미노산 서열을 외국에서 분리된 IHNV들과 비교 분석한 결과 90-95%의 상동성을 보였다. 이러한 사실은 INHV의 NV단백질 유전자들은 IHNV의 strain에 관계없이 매우 보존되어 있음을 나타내준다. Northern blotting을 사용하여 NV의 발현을 측정한 결과 감염 후 20 시간분터 발현이 증가함을 확인 힐수 있었다. NV가 바이러스의 증식에 필요한지의 여부를 확인하기 위하여 바이러스 유전자의 antisense DNA를 사용하여 바이러스 증식 억제에 관한 실험을 수행하였다. Glycoprotein (G)의 antisense DNA를 처리한 경우 바이러스의 증식이 거의 억제된 반면 NV에 대한 antisense DNA를 처리한 경우 바이러스 증식에 거의 변화가 없었다. 이로부터 배양중인 세포가 있어서 NV는 증식에 필수적이지 않은 것으로 판단된다. We have cloned and analyzed cDNA coding for non-virion (NV) protein of the m V - P R T The NV gene contained 336 bp open readmg frame and encoded a protein of 11 1 amino acids with a molecular weight of 13.2 kDa. The deduced amino acid sequence of NV of IHNVPRT was found to be 90-95% identical to those of foreign isolates of IHNV. These results indicate that NV gene of the MNV is highly conserved among &ifferent strains of THNV Northern blot analyses revealed that the levels of NV gene expression were strongly elevated after 20 h post-infection. In order to identify the role of NV in the replication of MNV in fish cells, IHNVinfected cells were treated with antisense oligonucleotides. While IHNV-PRT exposed to glycoprotein (G) antisense oligonucleotide showed severely reduced growth, the growth of virus exposed to NV antisense oligonucleotide was not affected by NV antisense oligonucleotide, which suggests that NV is not essential for replication of IHNV in fish cells.
이지연 ( Ji Yeon Lee ),조화자 ( Wha Ja Cho ),홍기성 ( Ki Sung Hong ),김계성 ( Kye Seong Kim ) 한국조직공학과 재생의학회 2004 조직공학과 재생의학 Vol.1 No.2
MicroRNAs (miRNAs) are family of small, non-coding RNAs that regulate gene expression in a sequence-specific manner. The two founding members of the miRNA family were originally identified in Caenorhbditis elegansas genes that were required for the timed regulation of developmental events. Since then, hundreds of miRNAs have been identified in almost all metazoan genomes, including worms, flies, plants and mammals. We identified expression of the miR-129b, miR-201, and miR-202 in various mouse tissues and development testis by Northern blot analysis. These three miRNAs have been registered in miRNA resistry as a testis specific miRNAs, however, only miR-129b was expressed in 2 weeks old mouse testis and also in spleen. miR-201 and miR-202 could not detected in not only testis but also other tissues.
한국에서 분리된 전염성 조혈괴저바이러스의 M 단백질의 유전자 클로닝과 염기서열 분석
박정민,김현주,문창훈,조화자,차승주,윤원준,박정재,이은희,박명애,손상규,박정우,Park, Jeong-Min,Kim, Hyun-Ju,Mun, Chang-Hoon,Cho, Wha-Ja,Cha, Seung-Ju,Yoon, Won-Joon,Park, Jeoug-Jae,Lee, Eun-Hee,Park, Myoung-Ae,Sohn, Sang-Gyu,Park, Jeon 한국미생물학회 1998 미생물학회지 Vol.34 No.1
한국에서 분리된 전염성 조혈괴저바이러스(infectious hematopoietic necrosis virus, IHNV)인 IHNV-PRT의 특성을 규명하기 위하여 IHNV-PRT의 matrix 단백질인 M1 및 M2를 암호화하고 있는 cDNA를 클로닝하여 이들의 염기서열을 분석하였다. M1은 693 bp 크기의 open reading frame을 포함하였으며 이로부터 230개의 아미노산으로 구성된 25.9 kDa의 분자량을 가진 단백질이 합성될 수 있다. M2는 588 bp 크기의 open reading frame을 지니고 있으며 195개의 아미노산으로 구성된 21.9 kDa의 분자량을 지닌 단백질이 합성될 수 있다. IHNV-PRT의 M1과 M2의 아미노산 서열을 외국에서 분리된 IHNV들과 비교 분석한 결과 M1은 92-93%, M2는 97%의 상동성을 보였다. 이러한 사실은 IHNV의 M 단백질 유전자들은 IHNV의 strain에 관계없이 매우 보존되어 있음을 나타내준다. In order to identify the characteristics of a Korean isol ate of infectious hematopoietic necrosis virus(IHNV), IHNV-PRT, we have cloned and analyzed cDNAs coding for matrix protein M1 and M2 of the IHNV-PRT. The Ml gene contained 693 bp open reading frame and encoded a protein of 230 amino acids with a molecular weight of 25.9 kDa. The M2 gene had 588 bp open reading frame, encoding a protein of 195 amino acids with a molecular weight of 21.9 kDa. On the deduced amino-acid sequences, M1 and M2 of the IHNV-PRT were found to be 92-93% (M1) and 97% (M2) identical to those of foreign isolates of IHNV. These results indicate that M genes of the IHNV are highly conserved among different strains of IHNV.
한국에서 분리된 전염성 조혈괴저바이러스의 N 단백질의 유전자 클로닝과 염기서열 분석
문창훈,김현주,박정민,조화자,차승주,윤원준,박정재,이은희,강호성,김한도,박정우,Mun, Chang-Hoon,Kim, Hyun-Ju,Park, Jeong-Min,Cho, Wha-Ja,Cha, Seung-Ju,Yoon, Won-Joon,Park, Jeong-Jae,Lee, Eun-Hee,Kang, Hoe-Sung,Kim, Han-Do,Park, Jeong-Wo 한국미생물학회 1998 미생물학회지 Vol.34 No.1
한국에서 분리된 전염성 조혈괴저바이러스(infectious hematopoietic necrosis virus, IHNV)인 IHNV-PRT의 nucleocapsid(N)를 암호화하고 있는 cDNA를 클로닝하여 분석하였다. N 유전자는 391개의 아미노산으로 구성된 42.3 kDa의 분자량을 가진 단백질을 암호화하는 1,176 bp 크기의 open reading frame을 가지고 있었다. N 단백질의 아미노산 서열을 다른 외국에서 분리한 IHNV들의 N과 비교한 결과 75-90% 정도의 유사성을 보였으나 다른 종의 fish rhabdovirus인 hirame rhabdovirus(HRV) 및 viral hemorrhagic septicemia virus(VHSV)와는 각각 43%와 38%의 유사성을 보였다. 그러나 아미노산 서열 214-265의 52개의 아미노산들은 모든 종의 fish rhabdovirus간에 매우 높은 유사성을 보여 주었다. We have cloned and analyzed cDNA coding for nucleocapsid protein N from infectious hematopoietic necrosis virus(IHNV), IHNV-PRT, which was isolated in Korea. The N gene had open reading frame of 1,176 bp that encoded a 391 amino acids with a molecular weight of 42.3 kDa. The deduced amino acid sequence of N protein was 75-90% identical to those of foreign isolates, IHNV, but was 43% and 38% identical to those of other species of fish rhabdovirus, hirame rhabdovirus(HRV) and viral hemorrahagic septicemia virus(VHSV), respectively. However, it revealed high levels of sequence identity between 214-265 amino acid sequences among all species of fish rhabdovirus.
Tristetraprolin Down-Regulates IL-23 Expression in Colon Cancer Cells
이현희,박정우,양성수,Mai-Tram Vo,조화자,이병주,임선희,차희정,이상현 한국분자세포생물학회 2013 Molecules and cells Vol.36 No.6
Interleukin 23 (IL-23) is an inflammatory cytokine that plays an important role in tumor promotion. Expression of IL-23 is increased in cancer cells and correlates with tumor progression. However, the mechanisms regulating IL-23 expression in cancer cells are still unclear. Here we report that tristetraprolin (TTP), an AU-rich element (ARE)-binding protein, inhibits IL-23 production in CT26 mouse colon cancer cells. Overexpression of TTP decreased the stability of IL-23 mRNA and the expression level of IL-23 in CT26 cells. Conversely, inhibition of TTP by siRNA increased IL-23 production. TTP destabilized a luciferase mRNA report-er containing the IL-23 mRNA 3’UTR, which contains five AREs. Analyses of deletion and point mutants of the IL-23 mRNA 3’UTR demonstrated that the ARE cluster between the third and fifth AREs was responsible for TTP-mediated destabilization of IL-23 mRNA. A RNA electrophoretic mobility shift assay confirmed that TTP binds to this ARE cluster. Taken together, these results demonstrate that TTP acts as a negative regulator of IL-23 gene expression in mouse colon cancer cells and suggest its potential appli-cation as a novel therapeutic target to control IL-23-mediated tumor promotion.