A comparative Study on the Integration of Exogenous DNA into Mouse, Rat, Rabbit, and Pig Genomes
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概要
- 論文の詳細を見る
Transgenic mammals, from small laboratory rodents to domestic animals, have been successfully produced to date, but their production efficiency within or across species has been variable. This is probably due to the differences in the type of injected DNA and/or technical procedures employed in each laboratory, as well as the reproductive characteristics of the species. Here we report the direct comparison of the efficiencies of producing transgenic mice, rats, rabbits and pigs by one technician using a fusion gene composed of the bovine αS1-casein promoter and human growth hormone (hGH) gene. Before the fusion gene was injected into the zygotes, high magnitude centrifugation to visualize the pronuclei was necessary for all of the pig zygotes and one-third of the rabbit zygotes, but not for mouse and rat zygotes. Post-injection survival of the mouse zygotes (67.1%) was lower than those of the rat, rabbit and pig zygotes (89.6 to 100%). The volume change of the pronucleus following DNA injection was the lowest in mice (50% increase), moderate in rabbits (148% increase), and the most prominent in rats (238% increase). The data from only 1 pig zygote indicated a 22% increase in the pronucleus volume by DNA injection. The PCR analyses of the tail DNA of new born offspring indicated that 0.8% (4/493), 4.8% (22/463), 0.8% (3/367) and 0.9% (2/221) of the injected eggs in mice, rats, rabbits and pigs, respectively, developed into transgenic offspring. Some of the founder animals in all four species expressed the transgene in the mammary gland which was confirmed in hGH mRNA by RT-PCR and/or hGH peptide in Witchs milk with ELISA. These results suggest that the maximum volume of DNA solution injectable into the pronucleus is a possible factor explaining the species differences in the production of transgenic animals.
- 社団法人 日本実験動物学会の論文
著者
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HIRABAYASHI Masumi
YS New Technology Institute, Inc.
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HOCHI Shinichi
Faculty of Textile Science and Technology, Shinshu University
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Ueda Masatsugu
YS New Technology Institute Inc., Shimotsuga-gun, Tochigi 329-0512, Japan
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ITO Kazumi
YS New Technology Institute, Inc.
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Ueda M
The Ys Institute Inc.
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Ueda Masatsugu
The Ys Inst. Inc.
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Ueda Masatsugu
ワイエスニューテクノロジー研究所
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HIRABAYASHI Masumi
National Institute for Physiological Sciences
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Hirabayashi M
National Inst. For Physiological Sciences
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Hirabayashi Masumi
自然科学研究機構生理学研究所
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Hirabayashi Masumi
信州大学 繊維
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Ito K
Ys New Technology Inst. Inc.
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Hochi S
Laboratory Of Theriogenology Obihiro University Of Agriculture And Veterinary Medicine
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Hochi Shinichi
Faculty Of Textile Science And Technology Shinshu University
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TAKAHASHI Ri-ichi
YS New Technology Institute. Inc.
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KASHIWAZAKI Naomi
YS New Technology Inst., Inc.
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HIRAO Masao
Ina Research Laboratory, Kitayama Labes Co. Ltd.
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HIRASAWA Kazuo
Ina Research Laboratory, Kitayama Labes Co. Ltd.
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Hirao Masao
Ina Research Laboratory Kitayama Labes Co. Ltd.
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Hirasawa Kazuo
Ina Research Laboratory Kitayama Labes Co. Ltd.
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Takahashi R
The Ys Institute Inc.
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Takahashi Ri-ichi
Ys New Technology Institute Inc.
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Ueda Masatsugu
Ys New Technology Institute
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TAKAHASHI Ri-ichi
The YS Institute Inc.
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Hirosawa Kensuke
Department Of Biomedical Science Faculty Of Agriculture The University Of Tokyo
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Kashiwazaki Naomi
Laboratory Of Animal Reproduction Azabu University
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Kashiwazaki N
School Of Veterinary Medicine Azabu University
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Kashiwazaki Naomi
Laboratory Of Animal Reproduction Graduate School Of Veterinary Science Azabu University
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Hochi S
Faculty Of Textile Science And Technology Shinshu University
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Ito Kazumi
Ys New Technology Institute
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Hirabayashi Masumi
Ys New Technology Institute Inc.
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Ueda Masatsugu
The Ys Institute Inc.
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Ueda Masatsugu
Ys New Tchnology Inst. Inc.
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Ito Kazumi
YS New Technology Inst., Inc.
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