Abstract
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.
Original language | English |
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Pages (from-to) | 213-216 |
Number of pages | 4 |
Journal | Asian Journal of Andrology |
Volume | 5 |
Issue number | 3 |
Publication status | Published - Sep 1 2003 |
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All Science Journal Classification (ASJC) codes
- Urology
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Evaluation of deoxyribonuclease activity in seminal plasma of ejaculated chicken semen. / Sato, Fuminori; Soh, Tomoki; Hattori, Masa Aki; Fujihara, Noboru.
In: Asian Journal of Andrology, Vol. 5, No. 3, 01.09.2003, p. 213-216.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Evaluation of deoxyribonuclease activity in seminal plasma of ejaculated chicken semen
AU - Sato, Fuminori
AU - Soh, Tomoki
AU - Hattori, Masa Aki
AU - Fujihara, Noboru
PY - 2003/9/1
Y1 - 2003/9/1
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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M3 - Article
C2 - 12937804
AN - SCOPUS:0042880031
VL - 5
SP - 213
EP - 216
JO - Asian Journal of Andrology
JF - Asian Journal of Andrology
SN - 1008-682X
IS - 3
ER -