Transgenic animal have extensive perspective of research and application in field of medicine, life science and biopharmacy. For this reason, scientists have been explored a more convenient and easy, economic and practical, and high-performance transgenic methods. Intratesticular injection as a method to transfer foreign gene into sperm can avoid inhibitory factor and complicated dealing of sperm, ensuring fertilization and embryo development. Recombinant efficient expression was made from pIRES2-EGFP plasmid backbone by inserting antimicrobial peptide -thanatin sequence constructed into the MCS region. Then the linear vector was injected into the testis of the BaiLai-hang chicken by surgical operation. 268 s were collected, 257 of them were transgenic chicken embryos. PCR and Southern blotting results revealed that 160 of them were positive transgenic chicken embryos, and transgenic rate was 59.70%. Southern blotting results showed that the foreign gene was integrated in chicken chromosomes with multi-inserted repeat sequences.