Abstract: A recombinant expression plasmid, plxas-N, was designed to produce antisense RNA targeted to the N gene full length of Transmissible gastroenteritis virus of swine, TGEV. The plxas-N was transfected into packaging cells line PA317 with Lipofectamine. The viral supenatants of the clones selected with G418 (500μg /ml) were detected by murine fibre cell NIH3T3.The highest viral titer pseudovirus was used to infect IBRS2 cell. Total cellular RNA from the IBRS2 cell, and RT-PCR analysis indicated that the plxas-N had inserted into the genome of IBRS2 cell. Antiviral activitity of the antisense RNA was evaluated by cytopathic effect using cultured IBRS2 cell and anti- IBRS2 cell infected by TGEV respectively, and the inhibitory rate was about 50 %.