| Co-Investigator(Kenkyū-buntansha) |
DATE Atsuko (ITO Atsuko) Ochanomizu Univ., Grad. Sch. of Hum. and Sci., Research Associate, 大学院・人間文化, 助手 (50303003)
KONDO Rumi Ochanomizu Univ., Dept. of Biolog, Assistant Professor, 理学部, 助手 (40293104)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Research Abstract |
In the previous study, we showed that Andropin has a rapid evolutionary pattern and amino acid sequences are not conserved between closely-related species. If a some kind of positive selection operates the Andropin evolution, it is considered that its function might slightly differ from each species. In this study, we tried to have a series of functional analysis ofAndropin in order to compare the gene expression pattern and protein activity between species.
First, we determined the sequence of 51 upstream regulatory region of Andropin between melanogaster closely-related species. It is not clear which transcription factor binding motif is necessary for tissue and stage-specific expression of Andropin. However, predicted transcription factor binding motifs are well conserved between the examined species. Interestingly, in these region, we found two TCTAA motifs, which may have a regulatory role of the tissue-specific expression pattern in the Drosophila male ejaculatory duct, referring
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the case of Glucose dehydrogenase (Gld) gene. This motif is also conserved between species. It may be considered that the expression pattern of Andropin is not different between species and that the diversification of Andropin from ancestral gene is accelerated by acquirement of such a motif in the regulatory region.
Second, we examined the expression patterns of Andropin between species by RT-PCR analysis. It is dear that Andropin gene expressions are shown among all of the melanogaster closely-related species, restricted in the adult male reproductive organ. In adult female, Andropin expression was not observed.
Next, in order to examine whether Andropin conserved its function among species, an antibacterial activity assay was performed. Andropin peptides were artificially synthesized for melanogaster and teissieri D. teissieri Andropin amino acid sequence shows the lowest amino acid identity with D. melanogaster Andropin and the predicted secondary structure was different from each other Assay was performed with the liquid method using three kinds of gram-positive bacteria and two of gram-negative bacteria. It has been already reported that D. melanogaster Andropin Mils not gram-negative bacteria, but gram-positive bacteria. As a result, antibacterial specificity of Andropin is not different between species ; active for gram-positive bacteria and not active for gram-negative bacteria. Although amino acid sequences vary considerably, it is considered that Andropin keeps its antibacterial activity between species in which have been examined Notably, hi the case of Bacillus subtilis, D. teissieri Andropin exerted its antibacterial activity in lower concentration than D. melanogaster Andropin. Less
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