Histopathological Changes of Liver, Lung and Kid- Ney in Ascites Syndrome in Broiler Chickens . Skeletal Disorders in Broiler Chickens (ross Breed) Estimation of Age In

s of papers 49 and still be immunogenic. In this study, our aim is to construct a bivalent recombinant ILTV with S1 gene inserted into the TK gene region of ILTV genome, The bivalent recombinant ILTV will express S1 gene and one of the potential application is a bivalent recombinant DNA vaccine for IBV and ILTV. Firstly, TK gene and S1 gene from ILTV and IBV was isolated with PCR or RT-PCR respectively. TK gene was then inserted into the Hind III/Pst I site of pBK-RSV phagemid, designated the recombinant as pBK-TK, secondly. S1 gene was then inserted into the SalI/ EcoR I site of pBK-TK, resulting recombinant is called pBK-TK-S1 and confirmed with DNA sequencing. After that, linearized pBK-TK-S1 recombinant plasmid was transfected with the Chicken embryo kidney cell (CEK) which had already been infected with a virulent ILTV Beijing E2 strain 6hrs before using a liposome transfection kit. Transfection mixture was then incubated at 370C in 5% CO2 in a humidified incubator for 5 days. The culture was frozen, thawed 3 times, and was passed it for two more times in CEK and incubated for the development of CPE. Viral DNA was prepared from transfection dishes with CPE was analyzed by PCR for the presence of the S1 gene. Positive samples were plaque purified and tested by PCR and immunofluoresence staining for the presence of the S1 gene. In conclusion, our result indicates a recombinant ILTV with S1gene of IBV inserted into the ILTV genome has been successfully constructed, and efforts are under way to investigate the immune efficacy of this recombinant ILTV in chicken.