4.1 Article

Generation of transgenic chicks using an oviduct-specific expression system

Journal

GENETICS AND MOLECULAR RESEARCH
Volume 10, Issue 4, Pages 3046-3055

Publisher

FUNPEC-EDITORA
DOI: 10.4238/2011.December.8.1

Keywords

Ovalbumin gene; GFP; Tissue plasminogen activator; Laying hen

Funding

  1. National High-Technology Research and Development Program of China (883) [2007AA100504]
  2. Annhui Natural Science Foundation [050410201]

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We successfully replaced the ovalbumin gene of a magnum region in chickens with a human plasminogen activator. We constructed pL-eGFP, pL-tPAGFP and pL-2.8OVtPAGFP vectors and cultured 293FT chicken embryo fibroblasts, chicken primordial germ cells, Hela C127 cells, and oviduct epithelial cells. All vectors were expressed in the transfected cells, except pL-2.8OVtPAGFP vector, which was only expressed in oviduct epithelial cells. A lentivirus with pL-2.8OVtPAGFP was injected in fertilized eggs; 11 chicks hatched in the G(0) generation, four of them carried the tPAGFP. Two cockerels from the G(0) generation were crossed with four wild-type hens. Three chicks in G(1) carried the tPAGFP. We concluded that by using an oviduct-specific vector for transfection, human recombinant plasminogen activator protein can be expressed in the oviducts of laying hens. This character is inherited and can be reproduced with a need for repeated transfection.

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