| Budget Amount *help |
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2003: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
|
|---|
| Research Abstract |
It takes long time and much cost to analyze the expressions of all known-functional genes by the conventional method. Furthermore, we cannot expect to utilize any commercial-based microarrays for oral bacterial in the future. Therefore, we prepared a DNA custom-made array of pathogenicity-associated genes in Porphyromonas gingivalis, strongly associated with disease activity in adult periodontal disease, and established the microarray procedure such as hybridization, wash, and labeling condition, and how to analyze. Consequently, mRNA was good start material for catching low signal intensities of gene expressions, compares with Total RNA. Moreover, it became clear to match results between microarray and Real-time PCR when we used fixed quantity of the labeled external standards from human genes for chip-chip normalization rather than 16S rRNA (internal standard). Following various environmental change was stressed to P. gingivalis and the transcriptome profiling was analyzed (1. Gene expression profiling during growth; 2. Gene expression profiling under hemin limitation; 3. Gene expression profiling under oxidization stress). These results suggested that we can expect not only to detect gene expressions of various pathogen but also to demonstrate the relationship among the pathogenic genes in P. gingivalis. Moreover, this technology expected a possibility that it could monitor of the degree of activity of the bacteria in a periodontal pocket.
|
