title: Testis mediated gene transfer: in vitro transfection in goat testis by electroporation.
abstract: Testis mediated gene transfer (TMGT) is a potential tool for making transgenic mice having more than 90% success rate. However, this method needs further standardization before it can be adapted in other species including livestock. In order to standardize the TMGT in goat, buck testes (n=20) collected from the slaughter house were injected with a vector driving green fluorescent protein (GFP) expression under a cytomegalovirus (CMV) promoter. Then, the testes were subjected to electroporation with predetermined voltage, pulse length, pulse interval and number of pulses. Seminiferous tubules were isolated from the electroporated testis and cultured in-vitro. The expression was checked at regular intervals. Green fluorescence was observed on different days in different samples. It suggests transient integration of the plasmid into the seminiferous tubules. This in-vitro transfection of seminiferous tubule using electroporation will provide valuable baseline information.
authors.affiliation: Division of Animal Genetics, Indian Veterinary Research Institute, Izatnagar, Bareilly Pin Code 243122, India. Electronic address: atishrraina@gmail.com.
authors.forename: Atish
authors.initials: A
authors.lastname: Raina
authors.affiliation: Division of Animal Genetics, Indian Veterinary Research Institute, Izatnagar, Bareilly Pin Code 243122, India.
authors.forename: Subodh
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Division of Animal Genetics, Indian Veterinary Research Institute, Izatnagar, Bareilly Pin Code 243122, India.
authors.forename: Rohit
authors.initials: R
authors.lastname: Shrivastava
authors.affiliation: Division of Animal Genetics, Indian Veterinary Research Institute, Izatnagar, Bareilly Pin Code 243122, India.
authors.forename: Abhijit
authors.initials: A
authors.lastname: Mitra
mesh_terms: D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics; D018274:Electroporation / Q000379:methods*; D018014:Gene Transfer Techniques; D006041:Goats; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008297:Male; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012671:Seminiferous Tubules / Q000378:metabolism*; D013737:Testis / Q000378:metabolism*; D014162:Transfection
chemical_list: D049452:Green Fluorescent Proteins
keywords: Electroporation; Seminiferous tubules; Testis mediated gene transfer; Transfection