The Study Of Technology Route Of Transgenic Chicken Producing With Liposome-mediated Green Fluorescent Protein Gene

The potential of using transgenic animals for the synthesis of therapeutic proteins was suggested over twenty years ago. Considerable progress has been made in developing methods for the production of transgenic animals and specifically in the expression of therapeutic proteins in the mammary glands of cows, sheep and goats. Increased demand for the production of human biopharmaceuticals in transgenic organisms has led to an intensive effort to develop the hen as a bioreactor producing exogenous proteins in egg white via transgenesis. To date, however, robust methods for transgenic modification of the avian genome have lagged behind in mammalian systems. The positive features associated with the use of the chicken in terms of cost, speed of development of a production flock and potentially appropriate glycosylation of target proteins have led to significant advances in transgenic chicken models in the past few years. So we carry out this rearch.Research Objective: This study was to transfer and express foreign genes under control of the ovalbumin promoter in the BCs. We have used plasmid DNA (pEGFP-C1) in combination with cationic liposomes to transfect chicken BCs. To obtain higher efficiency in liposome-mediated gene transfer into chicken BCs cells. A model was developed to explain the basis of anionic lipoplex uptake and transfection efficacy.Research Methods: The zygote laid by White Leghorn is used as expremental materials. In order to introduce exogenous DNA into gonads of chick embryos, stage X blastoderms of freshly laid and unincubated eggs were transfected by lipofection in vivo. Chicken BCs were isolated in vitro and cultured . Transfection efficiencies at different doses of DNA(1, 2, 3, 4, 5ug), liposome(6ul) were calculated under fluorescent microscopy using GFP(Green Fluorescent Protein) as a reporter gene. The liposome-DNA complex (1. 5ugDNA) were injected to the subgerminal cavity. We used microscopy and PCR to detect the GFP gene.