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KMID : 0900919990230010013
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Korean journal of Animal Reproduction
1999 Volume.23 No. 1 p.13 ~ p.18
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Production of Transgenic Animals by the Testis-Mediated Gene Transfer II. Production of Transgenic Korean Native Goats
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Yun Chang-Hyeon
Oh Seok-Doo
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Abstract
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The totipotential spermatogonial stem cells of adult testis which give rise to mature sperm cells is well-known to incorporate foreign DNA as well as those of somatic cells. Also, the integration rates of foreign DNA after haploid stages are generally known to decrease and /or is simply bound foreign DNA into the sperm plasma membrane. To overcome these problems, liposome and DNA complexes were used to determine how direct injection of these complexes into testis were integrated into sperm genome and resulted in transgenic offspring. To study this purpose, cation liposome was gently mixed with WAF/hGH DNA (1 : 2) and the complexes were injected into testis. At 10, 20, 40, 60, and 80 days after direct injection into testis, mature sperm cells were recovered by using artificial virgin method from two goats and each semen except a part of semen used for DNA analysis such as PCR or Southern blotting was cryopreserved for the artificial insemination. The results obtained in this study are summarized as follows. 1. By PCR, the presence of exogenous DNA was confirmed up to 80 days after injection with liposome/DNA complexes. The highest integration rates were obtained at day 40 after direct injection. This results suggested that spermatogonial stem cells were integrated exogenous DNA into their genome. 2. Among 23 Korean Native Goats which were artificially inseminated, 4 goats resulted in pregnancy and produced 7 young goats. 3. Two young goats were confirmed as a transgenic by PCR and Southern blot analysis. Therefore, our results suggested that testis-mediated gene transfer can be used as a feasible tools for the production of transgenic livestock.
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KEYWORD
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hGH, Spermatozoa, PCR, Korean Native Goat
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