| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2004: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
Based on the genomic information of Buchnera, the mechanism of the protein transport in an aphid endosymbiotic system was analyzed from the aspects of molecular biology and cellular biology, and following results were obtained.
1. It was found that the Buchnera 26 flagellar genes are not pseudogenes but all of them are actively transcribed and translated. Furthermore, electron microscopic observation revealed that the Buchnera surface is completely covered with thousands flagellum hook and basal body complexes (HBB). Analyses are planned to investigate the relation of the expression of flagellar genes and proteins to the developmental stages of host insects or morphogenesis.
2. In the course of the present study, we discovered that in alatae immediately after the final ecdysis, the number of Buchnera was decreased drastically, and also the hematoxylin-staining image of Buchnera was entirely different from that in apterae, suggesting that the localization of Buchnera DNA in alatae and apterae may be different.
3. As a preliminary experiment to prove the possibility that HBB functions as the protein transport machinery, isolated Buchnera were incubated with a fluorescent labeled substance and observed under a confocal microscope. Since the fluorescence was detected in Buchnera, construction of an in vitro assay system for protein transport was evaluated to be successful.
4. Several proteins which appeared to be imported from the host bacteria cells into Buchnera have been identified through proteome analysis. In the present investigation, it was the most important task to determine whether a "Buchnera targeting signal peptide" that is in common for these proteins exists or not. However, since the genome project for aphid is incomplete, our investigation has to be terminated. The results obtained in the present investigation will contribute in elucidating the mechanism of the protein transport mechanism, which is hitherto unknown, in a bacteriocyte endosymbiotic system.
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