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mRNA differential display 를 이용한 철에 의해 조절되는 유전자들의 분리 및 동정
이중림(Jung Lim Lee),박종환(Jong Hwan Park),김해영(Hae Yeong Kim) 한국응용생명화학회 1999 Applied Biological Chemistry (Appl Biol Chem) Vol.42 No.2
Iron is an essential nutrient but potentially toxic element in human. To identify the effects of iron on the gene expression of mammalian cell, we have isolated several genes that are regulated by iron using the RNA differential display method. RNAs were isolated from HeLa cells treated with iron supplement or iron chelator. A total of 24 genes were isolated and of these, four genes were identified by DNA sequencing and northern blot.
5'-Untranslated Region에 존재하는 Iron Responsive Element에 의한 Ferritin 합성조절
정인식,이중림,김해영,Chung, In-Sik,Lee, Jung-Lim,Kim, Hae-Yeong 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.33 No.2
철의 대사과정에 관여하는 ferritin 단백질의 발현은 ferritin transcript의 5'-untranslated region에 위치한 iron-responsive element (IRE)와 철 농도 조절 단백질의 결합에 의해 조절된다. 이러한 ferritin의 생성에 관여하는 구조적인 요소를 밝히기 위해, RNA 이차구조인 IRE의 bulge 부분을 다른 염기로 변환시켜 철 농도 조절단백질에 의한 RNA 결합력과 ferritin 단백질의 생성의 저해정도를 비교 측정하였다. 측정된 결과로부터 IRE의 bulge 부분의 시토신 염기배열만이 RNA 이차구조의 형성에 중요한 작용을 하여 ferritin 합성을 조절할 수 있는 것을 보였다. The expression of ferritin involved in iron metabolism is regulated at the translational level by the interaction of iron regulatory protein with iron-responsive element(IRE) in the 5'-untranslated region of ferritin transcript. To identify the role of structural element utilized for translational regulation of ferritin, we studied the effects of mutations in the ferritin IRE by measuring IRP binding activity and translational activity. Our data suggest that the cytosine at bulged position of IRE within ferritin is important for the formation of RNA secondary structure involved in translational regulation.
5'-Untranslated Region 에 존재하는 Iron Responsive Element 에 의한 Ferritin 합성조절
김해영,정인식,이중림 ( Hae Yeong Kim,In Sik Chung,Jung Lim Lee ) 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.41 No.3
The expression of ferritin involved in iron metabolism is regulated at the translational level by the interaction of iron regulatory protein with iron-responsive element(IRE) in the 5`-untranslated region of ferritin transcript. To identify the role of structural element utilized for translational regulation of ferritin, we studied the effects of mutations in the ferritin IRE by measuring IRP binding activity and translational activity. Our data suggest that the cytosine at bulged position of IRE within ferritin is important for the formation of RNA secondary structure involved in translational regulation.
이중림,김현중,김해영 경희대학교 생명자원과학연구원 1999 硏究論文集 Vol.20 No.-
사람 L-ferritin을 연구하기 위해 RT-PCR을 수행하여 ferritin 유전자를 T7 promoter expression vector에 subcloning시켜 대장균 내에서 발현시켰다 대량 발현된 L-ferritin의 발현 비율을 조사하였으며, SDS-PAGE 및, western blotting을 이용하여 재조합 I-ferritin의 발현을 확인하였다 Iron is an essential inorganic element for the growth of all forms of life. Iron is stored in cells in the form of ferritin. Human ferritins are composed of two types of subunits, Heavy(H)-and Light(L)-subunit. In this study, Human L-ferritin gene was isolated from human cell and subcloned into a bacterial expression vector. L-ferritin was overexpressed from E. coli BL21(DE3). By SDS-PAGE and western blotting, expressed protein was confirmed as feritin.
이중림,김현중,정인식,김해영 경희대학교 생명자원과학연구원 1998 遺傳工學論文集 Vol.10 No.-
사람 H-ferritin을 연구하기 위해 RT-PCR을 수행하여 ferritin 유전자를 polyhistidine fusion vector에 subcloning시켜 대장균내에서 발현시켰다. 대량발현된 H-ferritin은 nickel charged colum을 이용하여 ferritin을 정제하였다. Iron is an essential inorganic element for the growth of all forms of life. Iron is stored in cells in the form of ferritin. Human ferritins are composed of two types of subunits. Heavy (H). Light (L)-subunit. In this study, Human H-ferritin gene was subcloned into an expression vector pET15b maintained polyhistidine fusion system. H-ferritin was expressed and isolated from E. coli BL21 (DE3). By SDS-PAGE and western blotting. purified protein was confirmed as H-ferritin.
이중림,최동훈,김해영 경희대학교 생명자원과학연구원 1997 硏究論文集 Vol.18 No.-
Translation 과정에 영향을 미치는 RNA의 이차 구조와 구조의 존재하는 위치에 대한 영향을 조사하기 위해 본 연구에서는 model system으로 사람의 ferritin 유전자를 이용하였다. Ferritin 유전자의 5‘UTR, 단백질의 coding 지역내, 3’ UTR 등 3가지 위치에서 변형된 이차구조 인자를 이용하여 ferritin 단백질의 생성량을 in vitro translation을 이용하여 비교 측정하였다. 이 가운데 5‘ UTR에 존재하는 RNA 이차구조만이 ferritin 합성을 저해하였다. Genetic controls of cell metabolism and differentiation involve translational regulatory mechanism. Iron-responsive elements located within 5' or 3' untranslated regions(UTRs) are utilized for translational specific mRNAs. To identify the locations of structural elements utilized for translational regulation of mRNA, we modified and tested the roles of structural elements located within the 5', 3' UTR or coding region of ferritin. Our data suggest only 5'UTR element within ferritin is important for translational regulation.
최동훈,이중림,민승기,박종환,김해영,조재선 慶熙大學校 1998 論文集 Vol.27 No.-
Strain type of lactic acid bacteria was analysed by PCR and SDS-PAGE. PCR allows the rapid and specific detection of lactic acid bacteria using primers based on 16S rRNA gene-sequences. The analysis of protein pattern on SDS-PAGE can be used to determine the differences among the same or similar species of lactic acid bacteria. Therefore, the complementary use of PCR and SDS-PAGE method is one of simple way to classify the lactic acid bacteria and to compare with genetic analysis.