Generation of antiviral transgenic chicken using spermatogonial stem cell transfected in vivo

Abstract

This study was conducted in order to generate anti-viral transgenic chickens through transfected spermatogonial stem cell with fusion gene EGFP-MMx. After injecting fusion gene EGFP-MMx into testes, tissues frozen section, polymerase chain reaction (PCR) and dot blot of testes was performed at 30, 40, 50, 60, 70 and 80 days. Few normal hens were inseminated artificially with cock semen of 60 days. Finally, PCR, reverse transcription-PCR and Western blot were used to detect the integration of exogenous gene EGFP-MMx of the offspring. Dot blot positive rate of testes genome was 72.2%. PCR and RT-PCR positive rate was 25 and 10.53% (4/38) in sperm and F1 blood genome, respectively, while Western blot positive rate of F1 blood was 7.89% (3/38). Taken together, the possibility of cultivating anti-viral transgenic chickens by the method of testes injection which could integrate foreign genes into the genome was proved.

Key words: EGFP-MMx fusion protein, spermatogonial stem cell, transgenic chickens.