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- W2181393990 abstract "Sperm Mediated Gene Transfer (SMGT) is a technique based on the ability of sperm cells to bind exogenous DNA and can be used for the production of transgenic embryos and animals. The aim of this study was the immunolocation of the transgene bound to the spermatozoa after different sperm treatments (fresh-F, frozen-thawed-FT and quick frozen-QF) which induce different degrees of membrane alterations. Sperm samples were incubated with plasmid (EGFP, 5.4 kb) labelled by digoxigenin (DIG) in SFM medium (1x10 8 sperm/mL + 5µg DNA/mL). Anti-DIG labelled by FITC antibody was used for the localization of the DNA and observed by epifluorescent microscopy. Spermatozoa were classified in 3 categories according to DNA localization in the spermatozoa head: 1) in acrosomal region, 2) in post-acrosomal region, 3) in both regions (acrosomal and post-acrosomal regions). Using epifluorescent microscopy it was observed differences in the binding pattern after different sperm treatments, so in F sperm the DNA-binding is mainly localized in the acrosomal region (F: 51.09 ± 7.70%, FT: 17.77 ± 4.49%, QF: 7.76 ± 1.58%, P<0.01). On the other hand, when FT or QF spermatozoa were evaluated the DNA-binding tends to be in both regions (F: 40.04 ± 5.88%, FT: 54.82 ± 2.55%, QF: 78.09 ± 1.75%, P<0.01). In conclusion, there is a different exogenous DNA distribution in sperm depending on level of plasma membrane damage." @default.
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- W2181393990 date "2011-01-01" @default.
- W2181393990 modified "2023-09-26" @default.
- W2181393990 title "INMUNOLOCALIZACIÓN DE ADN EXÓGENO EN ESPERMATOZOIDES PORCINOS SOMETIDOS A DIFERENTES TRATAMIENTOS Immunolocation of Exogenous DNA in Boar Spermatozoa Under Different Treatments" @default.
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