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- 주제어 : Apin protein (검색결과 3 건)
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Ji-Woong Kim,Jong-Tae Park,You-Jin Lee,Hyun Sook Bae,Heung-Joong Kim,Kyoung-Yeon Lee,Seong-Ho Yun,JOO-CHEOL PARK 대한구강해부학회 2007 대한구강해부학회지 Vol.31 No.1
Apin protein, calcifying epitheli외 odontogenic (pindborg) tumors (CEOTs)-associated amyloid, were isolated from CEOTs, and has simi때 nucleotide sequences to OD314. It was reported that APin was highly expressed during ameloblast maturation and rnineralization. APin protein was 외 so efficiently secreted from cultured ameloblasts, However, little is known about the functional study of APin using recombinant protein during amelogeneis, ln this study, in order to produce the APin recombinat protein, the APin-expression construct were made and expressed its protein in various host cell and temperature conditions for the utilization of further enamel functional studies, The results were as follows: 1. APin protein was strongly expressed in the induction system using pRSET -OD314 construct 2. When the ]Mlæ was used as a expression host. APin protein was strongly expressed in the induction system using pHCEI田Nd-OD314 construct 3. The APin protein was recognized at a molecular weight of 22 kDa in Westem blots. 4. Almost of the expressed APin protein was soluble, These results suggest that considerable amount of APin recombinat protein was produced and it could be used for further amelogenesis research effectively.
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법랑모세포 분화와 법랑질 형성과정에서 OD314, Apin protein의 발현 및 기능
박종태,최용석,김흥중,정문진,오현주,신인철,박주철,손호현 대한치과보존학회 2006 Restorative Dentistry & Endodontics Vol.31 No.6
본 연구에서는 법랑모세포 분화와 법랑질 형성에 연관이 있는 OD314 일명 Apin protein의 기능을 밝힐 목적으로, in-situ hybridization에 의한 OD314 mRNA 발현과 법랑모세포 세포주에서 OD314 enamel matrix protein의 발현, 그리고 OD314 유전자를 과발현/억제시킬 수 있는 construct를 제작한 후 법랑질 형성 중에 OD314의 기능을 알아보고자 RT-PCR를 시행하여 다음과 같은 결과를 얻었다. 1. OD314 mRNA는 발생중인 상아모세포보다 법랑모세포에서 강하게 발현되었다. 2. Tuftelin은 석회화 결정이 형성되는 14일까지 발현이 지속되고, 그 이후부터 점차 감소하였다. Amelogenin과enamelin은 7일부터 그 발현이 점점 감소하였다. 3. U6-OD314 siRNA construct를 이용하여 transfection한 법랑모세포 세포주는 OD314와 tuftelin,MMP2 mRNA 발현이 감소하였으며, CM-OD314를 transfection하여 OD314의 과발현을 유도한 경우에는 OD314와 MMP20 mRNA의 발현이 뚜렷이 증대되었다. 이 결과는 OD314가 법랑모세포의 분화와 법랑질의 형성 그리고 석회화 과정에 중요한 역할을 하는 새로운 인자임을 시사한다. This study was aimed to elucidate the biological function of OD314 (Apin protein), which is related to ameloblast differentiation and amelogenesis. Apin protein, calcifying epithelial odontogenic (pindborg) tumors (CEOTs)-associated amyloid, were isolated from CEOTs, and has similar nucleotide sequences to OD314. We examined expression of the OD314 mRNA using in-situ hybridization during tooth development in mice. Expression of OD314 and several enamel matrix proteins were examined in the cultured ameloblast cell line up to 28 days by reverse transcription-polymerase chain reaction (RT-PCR) amplification. After inactivation and over-expression of the OD314 gene in ameloblast cell lines using U6 vector-driven RNA interference and CMV-OD314 construct, RT-PCR were performed to evaluate the effect of the OD314 during amelogenesis. The results were as follows: 1. In in-situ hybridization, OD314 mRNAs were more strongly expressed in ameloblast than odontoblast. 2. When ameloblast cells were cultured in the differentiation and mineralization medium for 28 days, the tuftelin mRNA expression was maintained from the beginning to day 14, and then gradually decreased to day 28. The expressions of amelogenin and enamelin were gradually decreased according to the ameloblast differentiation. 3. Inactivation of OD314 by U6-OD314 siRNA construct down-regulated the expression of OD314, MMP-20, and tuftelin, whereas over-expression of OD314 by CMV-OD314 construct up-regulated the expression of OD314 and MMP-20 without change in tuftelin. These results suggest that OD314 is considered as an ameloblast-enriched gene and may play the important roles in ameloblast differentiation and mineralization.
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