鸡减蛋综合征病毒河北分离株六邻体蛋白基因的克隆表达及免疫原性研究-预防兽医学专业论文.docxVIP

Cloning Cloning and Expression of Hexon gene from Egg Drop Syndrome Virus Detected in HeBei and Analysis of Immunogenicity Candidate：Du Donghua Specialty：Preventive Vetednary Medicine Advisor．Sun Jigun Abstract A strain of egg drop syndrome virus(EDSⅥwas detected from chicks which suffered from egg drop syndrome O!DS)in HeBei and provisionally named鹞HeBei strain(EDSV-HB)．A full length DNA clone for hexon gene ofthe vi鹏WaS constructed．and complete nuclcotide sequence ofthe gene was determined．And then the expression of the hexun-oncoding gene and the immunogenic of hexon protein were studied in this paper． According to the nucleotide sequence of egg drop syndrome virus AV-127 s11lin from the OeneBank,a pair of palmers specifically to amplify DNA of hexon gene of EDSV-HB SIFaln Was designed using Premer5．0 soflwme．PCR was performed for viral DNA extracted from the genome of EDSV-HB s缸ain using these primers and the amplified fragment Was inserted into pBS-T vector．These DNAs we他transformed into competent Esthetichis Coli TOPlO cells．Then the recombinantplasmid Was identified by digestion of restriction undonucleescs觚d PCR methods．The complete nucleotide sequence of the gene was determined．The result showed that the recombinant plasmid contained the complete sequence of the．hexon-encoding gane and the hexon-encoding gene was 2733 base pair long which encoded 910 amino acids,idcmtical with AV-127 strain and A^V-2 strain． The nucleotide and deduced amino acid sequences werc analyzed by computer program(DNAstar) using available data from data base(GeneBank)．Nucleotide homology and phylogenetic analysis were undertaken among HeBei strain and other strains of adenovims．Comparison witlI AV-127 strain indicated that there Wes 99．5％homology at the amino acids level．Comparison with^Av-2 sh面n showed that there was 98．4％homology al the amino acid level．which indicated that the hexon protein was conservative．Comparison ofnnclcotide sequence and deduced amino acid sequences ofhex