2006 Fiscal Year Final Research Report Summary
A novel RNA helicase-like protein during early embryonic development in silkworm Bombyx mori :
| Project/Area Number |
17580047
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Applied entomology
|
|---|
| Research Institution | Nihon University |
|---|
Principal Investigator |
SAWADA Hiroshi Nihon University, College of Humanities and Sciences, Associate Professor, 文理学部, 准教授 (60196326)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MASE Keisuke National Institute of Agrobiological Sciences Head of Sericultural Science Laboratory, Head of Sericultural Science Laboratory, 昆虫生産工学グループ・新蚕糸技術研究チーム, チーム長 (60414942)
YAMAHAMA Yumi Hamamatsu University, School of Medicine, Research associate, 医学部, 教務員 (90242784)
|
|---|
| Project Period (FY) |
2005 – 2006
|
| Keywords | DEAD-box / diapause / embryogenesis / recombinant protein / RNA helicase / silkworm / yolk cells |
|---|
| Research Abstract |
In order to understand the molecular mechanism of development during early embryogenesis in diapause and non-diapause of the silkworm, mRNA from diapause and non-diapause eggs was compared using the differential display technique. We cloned the full length of a cDNA encoding a novel RNA helicase-like protein (RHL) by the RACE method using a cDNA fragment which was one of the specific cDNAs in the non-diapause eggs. A BLAST search using the predicted amino acid sequence of RHL revealed a low homology (21-25% identity of its partial length) with that of the DEAD-box RNA helicase. Gene expression of the RHL gene of the diapause and non-diapause eggs was investigated by RT-PCR until 60 hr after oviposition. Amplified RHL cDNA was observed through all the stages in the non-diapause eggs, while in the diapause eggs, cDNA was found in eggs 0 to 12 hr after oviposition but disappeared 24 to 60 hr after oviposition. When the diapause eggs were activated by HCI treatment after chilling at 4 ℃ for 6 days from 48 hr after oviposition (artificial diapause termination), cDNA was observed from 12 hr after HCI treatment.
We also investigated the immunohistochemical distribution and localization of RHL in non-diapause eggs using anti-recombinant His-tag RHL antiserum. RHL was distributed in blastoderm cells and yolk cells and was localized in the nucleus and the cytosol of yolk cells. These data suggest that RHL has an important role in the early embryo of the silkworm.
|
