Cloning,Expression Of Structure Protein VP1 Of AEV And Preparation Of Monoclonal Antibodies Against VP1

Avian encephalomyelitis is an infectious disease of poultry which is characterized by neurological signs such as ataxia, paralysis and tremors of the neck and head.,Which can affect young chickens, pheasants, quail, and turkeys. This disease was of great economic importance to the poultry industry prior to the widespread use of vaccines. Now avian encephalomyelitis almost can be found in every country with poultry industry. It has been reported in some provinces of China since 1980, and is one of the serious diseases of poultry industry.It is very difficult for Avian Encephalomyelitis virus to be reproduced, which holds back the further research. Now, several new techniques such as indirect immunofluorescent assay (IFA) and indirect enzyme-linked immunosorbent assay (ELISA) have been developed to detect AEV, but both methods are cumbersome, time-consuming and expensive. Monoclonal antibodies are specific to antigens and considered a reliable method for diagnosis of pathogens.In order to clone the structure protein VP1 gene of AEV, and to develop special antibody against it, following work were finished to develop the plenty recombinant VP1 to detect the AEV in this experiment.1. Cloning and expression of structure protein VP1 gene of AEVBased on the published nucleotide sequence of VP1 gene of AEV strain 1143 , a pair of primers were designed and synthesized. Total RNA was isolated from primary purified AEV and used as template. The DNA fragments were amplified by RT-PCR and the result of nucleotide sequencing showed VP1 gene consists of 810 nucleotides and to...