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Exogenous DNA internalisation by sperm cells is improved by combining lipofection and restriction enzyme mediated integration.
- Source :
-
British poultry science [Br Poult Sci] 2011 Jun 01; Vol. 52 (3), pp. 287-91. - Publication Year :
- 2011
-
Abstract
- 1. Three types of exogenous DNA inserts, i.e. complete linearised pVIVO2-GFP/LacZ vector (9620 bp), the LacZ gene (5317 bp) and the GFP gene (2152 bp) were used to transfect chicken spermatozoa through simple incubation of sperm cells with insert. 2. PCR assay, Dot Blot hybridisation and Southern hybridisation showed the successful internalisation of exogenous DNA by chicken sperm cells. 3. Lipofection and Restriction Enzyme Mediated Integration (REMI) were used to improve the rate of internalisation of exogenous DNA by sperm cells. 4. Results from dot blot as well as Southern hybridisation were semi-quantified and improved exogenous DNA uptake by sperm cells through lipofection and REMI. Stronger signals were observed from hybridisation of LacZ as well as GFP specific probe with the DNA from lipofected exogenous DNA transfected sperm DNA in comparison with those transfected with nude exogenous DNA.
- Subjects :
- Animals
Blotting, Southern veterinary
DNA metabolism
DNA Restriction Enzymes metabolism
Green Fluorescent Proteins genetics
Green Fluorescent Proteins metabolism
Immunoblotting veterinary
Lac Operon
Liposomes
Male
Polymerase Chain Reaction veterinary
Transfection veterinary
Chickens genetics
DNA genetics
DNA Restriction Enzymes genetics
Gene Transfer Techniques veterinary
Spermatozoa
Transfection methods
Subjects
Details
- Language :
- English
- ISSN :
- 1466-1799
- Volume :
- 52
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- British poultry science
- Publication Type :
- Academic Journal
- Accession number :
- 21732873
- Full Text :
- https://doi.org/10.1080/00071668.2011.569927