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Introduction of foreign genes into animals forms the basis of a powerful approach for studying gene regulation. By using molecular biology methods genes of interest can be isolated, sequenced, recombined with regulatory elements and tested in vivo and in vitro for their functional ability. Chicken is a promising transgenic animal model for the production of therapeutically important biomolecules in egg for the pharmaceutical industry because of low feeding cost of hen, naturally sterile environment of egg, large amount of protein produced per egg and large number of eggs produced per hen. Production of transgenic chickens presents a set of problems since the manipulation of ovum like microinjection is difficult in birds because of their unique reproductive anatomy and physiology. Sperm mediated gene transfer offers a viable alternative for transgenic chicken production. The present study optimized conditions for efficient gene transfer in chicken sperms by nude gene transfer, lipofection and REMI. Internalization of foreign gene was verified by PCR assay and dot and Southern hybridization. A complete protocol for the production of transgenic spermatozoa in chicken was optimized.