1994 Fiscal Year Final Research Report Summary
Exploitation of new methods for efficent production of transgenic animals
| Project/Area Number |
05660323
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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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, Faculty of Agricclture, Associate Professor, 農学部, 助教授 (20041668)
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| Project Period (FY) |
1993 – 1994
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| Keywords | Transgenic / Bovine embryos / PCR / Farm animals |
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| Research Abstract |
Transgenic technology has provided powerful tools for the examination of the regulation of gene expression in cellular and physiological funtions. Transgenic farm animals havd been generated for improved products or productivity of domestic animals and resistance to animal diseases. (1) The most common method used for gene teansfere in farm animals is microinjection of DNA into pronuclei of zygotes. However, overall efficiency of transgenic farm animals were extreamely lower than that of miceowing to low integration of foreing DNA into host DNA.The present study was designed to improve the efficiency of generation of transgenic farm animals. (2) To improve the integration rate of for eign DNA into host DNA,the restriction enzyme with DNA was microinjected into the pronuclei of zygotes to induce the breaks in host genome DNA which increase the opportunity of integration of foreign DNA inothost DNA.In 8 concentrations from 10^<-4> to 10IU of EcoRI,The 10^<-1.5> and 10^<-2> IU were most effective to appropriate break in host genome DNA.To establish the method for selecting the transgenic embryos before they are transffered to the recipient, DpnI and Bal 31 enzymes were used for digestion of DNA fragement that not integrated into the host genome in the embryos. After this treatment, integrated gene was amplified by PCR.The results showed that this method was useful for selection of transgenic embryos. (3) As a basic experiment for methodological improvement of production rate of transgenic farm animals, the in vitro maturation, fertilization and culture of bovien oocytes cololected from the ovarian follicles of carcus were sutdied. It was found that individula differences in frozen semen remarkably influenced on the successful efficiency of in vitro fertilization.
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