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정소실질내 유전자 도입에 의한 형질전환동물의 생산 II. 형질전환 한국재래산양의 생산
윤창현,장규태,김성현,박미령,주학진,오석두,이병오 한국동물번식학회 1999 Reproductive & developmental biology Vol.23 No.1
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.
牛精液의 保存에 關한 硏究 : 原精液과 稀釋精液의 關係 Reration of Semen and diluted Semen
尹昌鉉 진주농과대학 1968 진주농과대학 연구논문집 Vol.- No.7
本試驗에 使用된 供試動物은 Holstein 2頭 Korean Cattle 2頭 計 4頭를 使用하였으며 試驗區는 稀釋保存液別로 3區로 設定하였다. 牛의 保存 精液에 있어서 原精液과 Seminan, Egg yolk citrate를 稀釋保存劑로서 使用한 精液을 保存하여 保存中의 精子生存指數를 調査함과 아울러 保存精液의 pH下降度를 調査하였던바 다음과 같은 結果를 얻었다. 1) 稀釋保存液間의 精子生存指數에 對하여 最少有意差를 檢定해 본 結果 T₁과 T₃間에는 5%水準의 有意差가 T₁과 T₂間에는 1%水準의 有意差가 認定되었다. 2) 牡牛間에는 有意性을 認定할 수 없었고 保存日間의 精子生存指數에 對해서는 1%水準의 高度有意性이 認定되었다. 3) 保存精液의 pH는 稀釋保存液間과 牡牛間에는 有意性을 認定할 수 없었고 保存日間에는 5%水準의 有意性을 認定할 수 있었다. 4) 精子의 生存指數 70% 以上을 使用可能限界라고 보면 T₂는 保存 6日에 限界에 達하고 T₃는 5日 T₁은 4日에 各各 限界線에 達하였다. 그러므로 Seminan의 成績이 가장 좋았고 다음이 Egg yolk, 原精液의 順位로 나타났다. 5) 保存日이 經過함에 따라 pH의 下降速度는 一致되지 않고 T₁이 가장 빨랐고 다음이 T₃T₂의 順位로 成績이 나타났다. 特히, T₂와 T₃의 下降速度가 느린것은 精液을 稀釋保存劑로서 5倍로 稀釋하여 保存精液內의 精子 濃度를 줄여 乳酸蓄積量을 줄인데 堤因된다고 思料된다. The effect of seminan and egg yolk-citrate diluter on the sperm motility and PH of bull semen. The experiment was conducted to study the effect of seminan and egg yolk-citrate diluter on the sperm motility and PH of bull seman collected from 2 Korean bulls and 2 Holstein bulls. The semen was preserved at 5℃ for 8 days and the test was reorded once a day through the period. The results obtained were as follows : 1. The sperm motility was singificantly higher(P<0.05) in the semen diluted with seminan than in the original semen, but not significantly different between bulls. 2. The sperm motility in all the treatments was decreased significantly as the time passed. 3. The average PH values between treatments and between bulls were found insignificantly different, but it was also decreased significantly (P<0.05) as the time passed. 4. The 70% sperm motility was maintained in the semen diluted with seminan, which could be considered as the lowest level to be inseminated successfully, in 6 days after treatment, but the lever was kept in the egg yolk-citrate diluter only for 5 days and in the original semen for 4 days after treatment. 5. The rapist decreasing rate of PH, as the time passed, was found in the original semen and the slowest in the semen diluted with egg yolk-citrate. It was considered that the slower decreasing rate found in the both diluted semen was due to the poor lactic acid accumulation by the less sperm concentration in the semen.
윤창현,박두순 한국산학기술학회 2003 한국산학기술학회 학술대회 Vol.- No.-
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