First study of sperm mediated gene transfer in Egyptian river buffalo

The present study was carried out to find the best treatments for enhancing the ration of insertion of a desired gene construct (pEGFP-N1) onto the sperm of buffalo as the first step for the production of transgenic buffalo using sperm mediated gene transfer (SMGT). The tested conditions were plasmi...

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Detalles Bibliográficos
Autores principales: Hassanane, Mohamed S., El Makawy, Aida I., Helalia, Sahar M., Abdoon, Ahmed S., Khalil, Kamal M.A., Ghanem, Tharwat A., Tohamy, Amany M., Sun, Xiao-Feng, Shen, Wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Academy of Scientific Research and Technology, Egypt 2017
Materias:
Animal Biotechnology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6296624/
https://www.ncbi.nlm.nih.gov/pubmed/30647689
http://dx.doi.org/10.1016/j.jgeb.2017.06.003
Descripción
Sumario:The present study was carried out to find the best treatments for enhancing the ration of insertion of a desired gene construct (pEGFP-N1) onto the sperm of buffalo as the first step for the production of transgenic buffalo using sperm mediated gene transfer (SMGT). The tested conditions were plasmid DNA concentration, sperm concentration, transfecting agent concentration: Dimethyle sulphoxide (DMSO) and time of transfection. The study proved that the best conditions for producing transgenic embryos were incubation sperm solution its concentration is 10(7)/ml sperm with 3% DMSO: with 20 µg/ml from the linarized DNA, for 15 min at 4 °C are the best conditions to produce transgenic buffalo embryo using sperm mediated gene transfer.

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