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Volume: 3, Issue: 4, July-August, 2015
Visible Diode Laser Enhancement of Exotic DNA Uptake by Fowl Sperm
Essam A. El-Gendy, Mona M. Abdelaziz, Mohamed M. Abdelfattah, Mohamed S. Salama, YhiaM. Badr
An experiment was conducted to assess the use of low power laser irradiation and lipofectin to enhance fowl sperm uptake of exogenous DNA. Semen samples of 10 roosters were collected and pooled. The pooled sample was diluted with a semen extender and then divided into 6 aliquots for 6 different treatments. The treatments included different combinations of the inclusion of exogenous DNA (bacterial plasmid pUC18), the exposure to low power laser irradiation and the transfection with lipofectin (5%). Laser irradiation was by using visible diode laser (650 nm) at energy dose of 4 J/cm2. The recognition of the plasmid DNA in the sperm was by using two specific oligonucleotides (forward and reverse) to prime a 420-bp fragment on the pMB1 rep of the plasmid. The results indicated that low power laser irradiation enhanced the sperm uptake of the plasmid DNA. Also, lipofectin enhanced the introduction of the plasmid DNAinto the sperm, whether the semen was laser irradiated or not.
Keywords: Fowl sperm, laser irradiation, lipofectin, exotic DNA.