TransgenicChickenswithoviductspecificvector.docVIP

Transgenic Chicken with Oviduct Specific Vector Abstract The present study was attempted to replace ovalbumin gene with tissue plasminogene activator marked with green fluorescence protein. The pl-EGFP, pl-CMV-tPAEGFP and pl-2.8OV-tPAEGFP vectors were constructed. 293FT cells were transfected with pl-EGFP vector and chicken embryo fibroblasts (CEF), chicken primary germ (cPGC) cells with pl-CMV-tPAEGFP vector. The green fluorescence protein was expressed among all trasnfected cells. The pl-2.8OV-t-PAGFP and control (pEGP-N1) vector was transfected with Hela, C127 and oviduct epithelial cells. The pl-2.8OV-t-PAGFP vector was only expressed in oviduct epithelial cells but not in hela and C127 while control vector was expressed in all hela, C127 and oviduct epithelial cells. The result showed that pl-EGFP, pl-CMV-tPAEGFP vectors could infect different cells but pl-2.8OV-t-PAGFP vector was only expressed in Oviduct epithelial cells due to oviduct specific expression system. The lentivirus with (pl-2.8OV-tPAEGFP) was injected in 50 fertilized eggs, 11 (22%) chicken hatched with 4 (36%) positive containing integration of exogenous genes analyzed by DNA dot blotting and PCR. Our finding suggested that transgenic chicken can be produced using oviduct specific vector containing pl-2.8OV-tPAEGFP. Key Word: Lentiviral vector; ovalbumin gene; tissue plasminogene activator; GFP; laying hens Introduction Presently one of the most advanced research of is genetically modified technology, it has been involved in production of transgenic animals. The ?rst transgenic animals were generated by infecting mouse blastocyst cells with a Moloney leukemia virus-based retroviral vector (Jaeisch 1988, 1976). Lentiviral vector has been used successfully to create transgenic mice with a very high rate of incorporation of the foreign gene (Lois et al., 2002). Lentiviral vec