| Project/Area Number |
11694226
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
蚕糸・昆虫利用学
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| Research Institution | National Institute of Radiological Sciences |
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Principal Investigator |
MITA Kazuei National Institute of Radiological Sciences, Genome Research Group, Researcher, 第2研究グループ, 研究員 (30159165)
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| Co-Investigator(Kenkyū-buntansha) |
ICHIMURA Sachiko National Institute of Radiological Sciences, Researcher, 生物影響研究部, 研究員 (40158754)
MORIMYO Mitsuoki National Institute of Radiological Sciences, Genome Research Group, Researcher, 第2研究グループ, 研究員
SHIMADA Toru University of Tokyo, Department of Agriculttural and Environmental Biology, Associate Professor, 農学生命科学研究科, 助教授 (20202111)
TAMURA Toshiki National Institute of Sericultural and Entomological Science, Researcher, 蚕糸・昆虫農業技研究所遺伝育種部, 研究員
OSOEGAWA Kazutoyo Roswell Park Center Institute, Genetic Department, Research Associate, Research Associate
森明 先興 放射線医学総合研究所, 第2研究グループ, 研究員 (00166430)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 2000: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1999: ¥3,200,000 (Direct Cost: ¥3,200,000)
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| Keywords | silkworm, Bombyx mori / BAC library / BAC contig / hybridization / EST / physical map / fingerprinting / フィンガープリント / BACコンティーグ / 遺伝子地図 / カイコゲノム解析 |
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| Research Abstract |
The purpose of the project is the construction of the physical map of silkworm genome for the silkworm genome analysis. First, we succeeded in the construction of a high quality BAC library. The BAC library contains approximately 36,000 clones with an average insert size of 168 kb, which corresponds to 11-fold genome redundancy. The rate of chimera was estimated to be less than 3%. The BAC clones were arrayed into 96 384-well microtiter dishes and were gridded in duplicate in specific patterns onto two nylon membranes (High Density Rplica (HDR) filters). We are employing two methods for the construction of BAC contigs covering whole genome, (1)the EST maker-overlapping method and (2)the fingerprinting method. For the overlapping of EST markers, BAC clones containing EST sequences were screened by hybridization of HDR filters using ESTs as probes. The genome size of silkworm is estimated to be 530 Mb. Since the average size of BAC clones is 168 kb, more than 6,200 hybridization experiments using non-redundant EST probes are required to complete BAC contig construction. So far, about 2,500 hybridizations were over. The fingerprinting method based on restriction enzyme cutting patterns is more effective than EST marker-overlapping method for the regions with low gene density, where the EST-overlapping method is not available. For fingerprinting experiments, about 1/3 experiments were carried out so far. The construction of whole BAC contigs using tose two methods are progressing. For several loci of Z and W sex chromosomes, chorion locus of chromosome 2 and virus resistant gene locus of chromosome 17, the BAC contigs were constructed and analyses using the BAC clones revealed characteristic features of silkworm genome and interesting genes.
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