title: Exogenous DNA internalisation by sperm cells is improved by combining lipofection and restriction enzyme mediated integration.
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.
authors: Genome Mapping Laboratory, Avian Genetics & Breeding, Central Avian Research Institute, Izatnagar, Uttar Pradesh, India.
authors: R R
authors: RR
authors: Churchil
authors: J
authors: J
authors: Gupta
authors: A
authors: A
authors: Singh
authors: D
authors: D
authors: Sharma
mesh_terms: D000818:Animals; D015139:Blotting, Southern / Q000662:veterinary; D002645:Chickens / Q000235:genetics*; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000235:genetics* / Q000378:metabolism; D018014:Gene Transfer Techniques / Q000662:veterinary*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D015151:Immunoblotting / Q000662:veterinary; D007763:Lac Operon; D008081:Liposomes; D008297:Male; D016133:Polymerase Chain Reaction / Q000662:veterinary; D013094:Spermatozoa; D014162:Transfection / Q000379:methods* / Q000662:veterinary
chemical_list: D008081:Liposomes; D049452:Green Fluorescent Proteins; D004247:DNA; D004262:DNA Restriction Enzymes