Evaluation of deoxyribonuclease activity in seminal plasma of ejaculated chicken semen / 亚洲男科学杂志(英文版)
Asian Journal of Andrology
;
(6): 213-216, 2003.
Article
in English
| WPRIM
| ID: wpr-300889
ABSTRACT
<p><b>AIM</b>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.</p><p><b>METHODS</b>A PCR fragment (249 bp) of pEGFPN-1 vector was used as the DNA substrate and was incubated with the seminal plasma at 40 degree C for 30 min. Then, the whole reaction solution was subjected to agarose gel electrophoresis and the DNA size was evaluated under UV light.</p><p><b>RESULTS</b>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 approximately 5 mmol/L EDTA. The substrate was stabilized in the seminal plasma by the addition of commercially available lipofection reagents.</p><p><b>CONCLUSION</b>The DNase activity is present in the seminal plasma of ejaculated chicken semen. However, DNA is stable in the liposomal-DNA complex.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Physiology
/
Semen
/
DNA
/
Cations, Divalent
/
Chickens
/
Edetic Acid
/
DNA Primers
/
Reverse Transcriptase Polymerase Chain Reaction
/
Deoxyribonucleases
Limits:
Animals
Language:
English
Journal:
Asian Journal of Andrology
Year:
2003
Type:
Article
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