1998 Fiscal Year Final Research Report Summary
Basic studies on production and application of transgenic animals in ruminants
| Project/Area Number |
08406018
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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 | 一般 |
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| Research Field |
Applied animal science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
TOJO Hideaki The University of Tokyo, Graduate School of Agricultural and Life Sciences, Professor, 大学院・農学生命科学研究科, 教授 (20041668)
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| Co-Investigator(Kenkyū-buntansha) |
YAMANOUCHI Keitaro The University of Tokyo, Graduate School of Agricultural and Life Sciences, Assi, 大学院・農学生命科学研究科, 助手 (70272440)
SAKAI Senkichi The University of Tokyo, Graduate School of Agricultural and Life Sciences, Prof, 大学院・農学生命科学研究科, 教授 (80114487)
SAWASAKI Tooru The University of Tokyo, Graduate School of Agricultural and Life Sciences, Prof, 大学院・農学生命科学研究科, 教授 (00012047)
TKAHASHI Michio The University of Tokyo, Graduate School of Agricultural and Life Sciences, Prof, 大学院・農学生命科学研究科, 教授 (30011943)
TACHI Chikashi Azabu University, School of Veterinary Sciences, Professor, 獣医学部, 教授 (30011711)
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| Project Period (FY) |
1996 – 1998
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| Keywords | transgenic / homeobox / c-kit / mouse / caprine hircus / EGFP |
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| Research Abstract |
1. The complete open reading tram of the c-kit cDNA was clone from the Shiba goat (Capra hircus var Shiba) with the dominant black-eyed white phenotype. The analysis of the deduced amino acid sequence revealed the presence of a single amino acid insertion (alanin) in the kinase insert region. 2. The expressions of homeobox genes in murine and caprine placentae were surveyed. In the murine placentae, all the Hox 9 paralogues except for Hox c9 were expressed. In the caprine placentae, the HOX 9 paralogues except for HOX d9 were detected. 3. Attempts were made to investigate whether co-injection of foreign DNA constructs with bacterial restriction endonuciease into the pronucleus of mouse zygotes will be capable of elevating integration frequency of foreign DNA into the host genome. The results showed that co-injection of the genes with restriction enzyme appeared the slight elevation of successful integration in the embryos and the pups. 4. One way to improve the efficiency of transgenic animal production would be to identify the transgenic embryos before transfer into foster mothers. Green fluorescent protein (GFP) can be a suitable marker for the selection of transgenic embryos. CMV beta-actin/EGFP gene was microinjected into the pronucleus of mouse zygotes. The DNA injected embryos were cultured in vitro and observed for fluorescence. The fluorescent embryos were transferred into the uteri of recipient mice. Pups born were analyzed for transgenic. A part of the embryos were analyzed for the transgene by the Dpn I-Bal31 digestion method. The results showed that approximately 77% of the embryos with uniform fluorescence were transgenic. The present study demonstrated that the method using EGEP as a marker be very reliable for selecting the transgenic embryos.
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