CONSTRUCTION AND APPLICATION OF CDNA MICROARRAY SPECIALIZED FOR PERIODONTAL IIGAMENT

1.人牙周膜基因表达谱分析利用人牙周膜(PDL)三向cDNA文库收集的序列，真实地反映了人牙周膜基因群体的组成。利用这些信息，我们研究了活性基因的表达谱。我们揭示了PDL具有独特的活性基因表达谱。新基因PLAP-1的鉴定和鉴定通过对人PDLα基因表达谱的分析，我们鉴定了一个新基因，它编码富含电影的小分子重复蛋白多糖家族的一个新成员。我们将其命名为PLAP-1(牙周膜相关蛋白-1)。研究表明，PLAP-1在体内的PDL中特异表达，在维持PDL.3的动态平衡中起重要作用。定制的人PDL基因芯片的构建选择在人PDL中表达的基因，我们构建了专门针对人PDL的基因芯片，…此外，我们还命名为PerioGen Chip PerioGen Chip，该芯片包含从人PDL.4的3-Directed cDNA文库中获得的1600个cDNA克隆。利用PerioGen芯片分析PDL的基因表达，分析PDL细胞在体外分化过程中的基因表达模式。PerioGen Chip分析发现，有15个基因在细胞分化过程中表现出两倍或更多的差异表达。在这些基因中，我们重点研究了对其转录具有很强诱导作用的DAN基因和Periostin基因。我们证明了DAN和Periostin对PDL细胞的细胞分化具有重要的调控作用。用PerioGen芯片进行活体牙周组织基因表达分析我们对牙周病患者牙周组织进行了基因表达分析。我们能够鉴定出几个主要在牙周病变中表达的基因。这表明PerioGen芯片可以用于临床应用。较少

1. Analysis of gene expression profile of human periodontal ligamentWe collected sequences with the 3 -directed cDNA library of human periodontal ligament (PDL), which faithfully represents the composition of the mRNA population. Utilizing the information, we investigated the expression profile of the active genes. We revealed that PDL has a unique active gene expression profile.2. Identification and characterization of a novel gene, PLAP-1By analyzing the a gene expression profile of human PDL, we identified a novel gene, which encodes a new member of the small let cine-rich repeat proteoglycan family. We designated it PLAP-1(periodontal ligament associated protein-1). It was shown that PLAP-1 is expressed specifically in PDL in vivo and plays an important role in maintaining homeostasis of PDL.3. Construction of customized cDNA microarray specialized for human PDLSelecting genes that are expressed in human PDL, we constructed in-house cDNA microarrays specialized for human PDL, which … More we named PerioGen Chip PerioGen Chip contained 1,600 cDNA clones obtained from the 3 -directed cDNA library of human PDL.4. Gene expression analysis of PDL utilizing PerioGen Chip Utilizing PerioGen Chipwe analyzed the patterns of gene expression during the cytodifferentiation of PDL cells in vitro. PerioGen Chip analysis identified 15 genes showing twofold or more differential expression during the cytodifferentiation. Among these genes, we focused on DAN gene and Periostin gene which showed strong induction of their transcript. We demonstrated that DAN and Periostin have crucial roles to regulate cytodifferentiation of PDL cells.5. Gene expression analysis of in vivo periodontal tissues by PerioGen ChipWe performed gene expression analysis of periodontal tissues which were derived from patients with periodontal diseases. We were able to identify several genes which were expressed predominantly in periodontal lesions. This demonstrated that PerioGen Chip can be useful for clinical application. Less