Evaluation of deoxyribonuclease activity in seminal plasma of ejaculated chicken semen

Fuminori Sato, Tomoki Soh, Masa Aki Hattori, Noboru Fujihara

研究成果: ジャーナルへの寄稿記事

10 引用 (Scopus)

抄録

Aim: To confirm the stability of exogenous genes in the generation of transgenic chickens using ejaculated chicken sperm, the deoxyribonuclease (DNase) activity was evaluated in the seminal plasma of ejaculated semen and the stability of DNA was examined by adding lipofection reagents. Methods: A PCR fragment (249 bp) of pEGFPN-1 vector was used as the DNA substrate and was incubated with the seminal plasma at 40 °C for 30 min. Then, the whole reaction solution was subjected to agarose gel electrophoresis and the DNA size was evaluated under UV light. Results: The DNA substrate was completely diminished after incubation with seminal plasma. However, the substrate was intact after incubation with heat-treated seminal plasma or incubation with seminal plasma in the presence of 0.5 mmol/L ∼5 mmol/L EDTA. The substrate was stabilized in the seminal plasma by the addition of commercially available lipofection reagents. Conclusion: The DNase activity is present in the seminal plasma of ejaculated chicken semen. However, DNA is stable in the liposomal-DNA complex.

元の言語英語
ページ(範囲)213-216
ページ数4
ジャーナルAsian Journal of Andrology
5
発行部数3
出版物ステータス出版済み - 9 1 2003

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Deoxyribonucleases
Semen
Chickens
DNA
Agar Gel Electrophoresis
Ultraviolet Rays
Edetic Acid
Spermatozoa
Hot Temperature
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Urology

これを引用

Evaluation of deoxyribonuclease activity in seminal plasma of ejaculated chicken semen. / Sato, Fuminori; Soh, Tomoki; Hattori, Masa Aki; Fujihara, Noboru.

:: Asian Journal of Andrology, 巻 5, 番号 3, 01.09.2003, p. 213-216.

研究成果: ジャーナルへの寄稿記事

Sato, Fuminori ; Soh, Tomoki ; Hattori, Masa Aki ; Fujihara, Noboru. / Evaluation of deoxyribonuclease activity in seminal plasma of ejaculated chicken semen. :: Asian Journal of Andrology. 2003 ; 巻 5, 番号 3. pp. 213-216.
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N2 - Aim: To confirm the stability of exogenous genes in the generation of transgenic chickens using ejaculated chicken sperm, the deoxyribonuclease (DNase) activity was evaluated in the seminal plasma of ejaculated semen and the stability of DNA was examined by adding lipofection reagents. Methods: A PCR fragment (249 bp) of pEGFPN-1 vector was used as the DNA substrate and was incubated with the seminal plasma at 40 °C for 30 min. Then, the whole reaction solution was subjected to agarose gel electrophoresis and the DNA size was evaluated under UV light. Results: The DNA substrate was completely diminished after incubation with seminal plasma. However, the substrate was intact after incubation with heat-treated seminal plasma or incubation with seminal plasma in the presence of 0.5 mmol/L ∼5 mmol/L EDTA. The substrate was stabilized in the seminal plasma by the addition of commercially available lipofection reagents. Conclusion: The DNase activity is present in the seminal plasma of ejaculated chicken semen. However, DNA is stable in the liposomal-DNA complex.

AB - Aim: To confirm the stability of exogenous genes in the generation of transgenic chickens using ejaculated chicken sperm, the deoxyribonuclease (DNase) activity was evaluated in the seminal plasma of ejaculated semen and the stability of DNA was examined by adding lipofection reagents. Methods: A PCR fragment (249 bp) of pEGFPN-1 vector was used as the DNA substrate and was incubated with the seminal plasma at 40 °C for 30 min. Then, the whole reaction solution was subjected to agarose gel electrophoresis and the DNA size was evaluated under UV light. Results: The DNA substrate was completely diminished after incubation with seminal plasma. However, the substrate was intact after incubation with heat-treated seminal plasma or incubation with seminal plasma in the presence of 0.5 mmol/L ∼5 mmol/L EDTA. The substrate was stabilized in the seminal plasma by the addition of commercially available lipofection reagents. Conclusion: The DNase activity is present in the seminal plasma of ejaculated chicken semen. However, DNA is stable in the liposomal-DNA complex.

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