| Project/Area Number |
07557205
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
寄生虫学(含医用動物学)
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| Research Institution | OBIHIRO UNIVERSITY OF AGRICULTURE AND VETERINARY MEDICINE |
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Principal Investigator |
AGATSUMA Takeshi OBIHIRO UNIVERSITY OF AGRICULTURE AND VETERINARY MEDICINE,DEPARTNENT OF BIORESOURSE SCIENCE,ASSOCIATE PROFESSOR, 畜産学部, 助教授 (40117031)
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| Co-Investigator(Kenkyū-buntansha) |
TAGUCHI Takahiro KOCHI MEDICAL SCHOOL,DEPARTMENT OF MEDICINE,INSTRUCTOR, 医学部, 助手 (80127943)
HIRAI Hirohisa KYOTO UNIVERSITY,DEPARTMENT OF MEDICINE,ASSISTANT PROFESSOR, 霊長類研究所, 助手 (10128308)
HIRATA Mizuki KURUME UNIVERSITY,DEPARTMENT OF MEDICINE,ASSOCIATE PROFESSOR, 医学部, 助教授 (70080629)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 1996: ¥5,800,000 (Direct Cost: ¥5,800,000)
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| Keywords | SCHISTOSOMA JAPONICUM / GENOME MAPPING / CHROMOSOME DISSECTION / FISH / PRINS / PAINTING PROBE / C-BANDING PATTERN / SNAIL HOST / 染色体 / DNA / ゲノム解析 / FISH / 染色体顕微切断 / PRINS |
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| Research Abstract |
In this project, we developed three items as following : (1) Chromosome microdissection and fish : chromosome preparation was made on a cover glass (24x60 mm^2) using cells fixed by ethanol (99.5%). The cells were cultured with RPMI 1640 included 20% FCS,3mug/ml LPS,and 2mug/ml conA.Chromosomes on the preparations were dissected by using a micro glass knife with 2mum diameter on an inverted microscope. The microdissected chromosome fragments were collected into 0.5ml tube with collection drop. PCR products obtained were localized by the fish techniques. (2) PCR procedure : microdissected fragments were transferred into 0.5ml microtube containing 1ul collection drop(2.5mm EDTA,1mg/ml proteinase K,41% polyethylene glycol 6,000). After deproteinization, the 1st PCR amplification was carried out using universal primer A (5-gga aac agc tat gac ctg aat tcn nnn nna tgt gg-3). The second PCR anplification was performed using primer B (5-gga aac agc tat gac ctg aat tc-3). PCR Iabeling was final
… More
ly done using the primer B and biotin. The labeled products were localized by fish to check their qualities. (3) Application of the chromosome microdissection techniclue : a) Chromosome microdissection : in S.japonicum, the 1st chromosome (both of short and long arms) , the 2nd chromosome (SJ2) and the ZW chromosome were dissected and tried to be localized by the FISH method. Only the SJ2 chromosome was hybridized with every telomer region of all of the chromosomes. For Robertsiella sp. (the intermediate host of S.malaynsis), the 1st and Y chromosomes, and, the 1st and 2nd chromosomes for Oncomelania hupensis (the intermediate host of S.japonicum) were tried to dissect and paint. Many strong signals were detected on the whole chromosomes in both host species. B) PRINS : three types were observed on the basis of painted regions. (1) every end of all the chromosomes : S.japonicum S.sinensium (2) every end of all the chromosomes, centromere and heterochromotin of the W chromosome : S.mansoni (3) every end of all the chromosomes and heterochromotin of the W chromosome Less
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