禽腺病毒W株Hexon基因的分子特征及其致鸡胚侏儒化研究  

Molecular characteristics of the Hexon gene of the fowl Adenvirus W strain and its dwarfization in embryonated chicken eggs

作　　者：陈田田 刘琳[1] 赵玉杰[1] 刘建勋 王赛楠 何春辉 李新生[1] CHEN Tiantian;LIU Lin;ZHAO Yujie;LIU Jianxun;WANG Sainan;HE Chunhui;LI Xinsheng(College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China)

机构地区：[1]河南农业大学牧医工程学院

出　　处：《畜牧与兽医》2019年第9期81-86,共6页Animal Husbandry & Veterinary Medicine

基　　金：国家十三五专项(2017YFD0500701-5);河南省科技开放合作基金(182106000048);河南省高校科技创新团队支持计划(19IRTSTHN007)

摘　　要：从河南焦作某发生心包积液和肝肿大为病变特征的病鸡群采集肝脏组织,经分离、纯化、PCR鉴定和测序分析,确定病原为I群C种血清4型禽腺病毒(FAdV-4),命名为W株。该W株的ELD50为10^-3.576/0.2 mL,Hexon基因全长2 814 bp,A、T、G、C碱基数和含量分别为654个(23.24%),514个(18.27%),679个(24.13%)和967个(34.36%)。Hexon基因编码蛋白的分子量为106.7 ku,共含有937个氨基酸,其中疏水氨基酸439个(46.80%),中性氨基酸265个(28.25%),亲水氨基酸233个(24.84%)。W株Hexon基因推导的氨基酸序列与FAdV-4 JSJ13株相比,在第181、939位2个氨基酸位点存在缺失;与FAdV-4 PK-01株相比,在第194、371、376位3个氨基酸位点存在突变;与FAdV-4 ON1株相比,有25个氨基酸位点存在突变。将W株经绒毛尿囊膜途径接种10日龄SPF鸡胚,与未接种病毒的对照组相比,接种W株病毒组在接种后第3天EE值均呈现显著性差异(P<0.05),第6天呈现极显著性差异(P<0.01),从而确定EE值和EE指数可以作为反映FAdV-4对鸡胚致侏儒作用的重要指标。In this study, the liver tissues were collected from the chickens infected with hydropericardium and hepatomegaly in Jiaozuo City, Henan Province.Isolation, purification, PCR and sequencing were used to identify the pathogen to be the fowl Adenvirus serotype 4(FAdV-4) in species C which was designated as the W strain(10^-3.576 ELD50/0.2 mL). The results showed that the Hexon gene of the W strain was 2814 bp in length, in which the number and content of bases A, T, G and C were: A, 654(23.24%);T, 514(18.27%);G, 679(24.13%);and C, 967(34.36%);and the average molecular weight of the amino acid sequence deduced by the Hexon gene was 106.7 ku, encoding 937 amino acids, of which 439 were hydrophobic amino acids(46.80%), 265 were neutral amino acids(28.25%), and 233 were hydrophilic amino acids(24.84%). The amino acid sequence deduced by the Hexon gene of the W strain showed 2 amino acids in deletion, compared with JSJ13 strain at the sites of 181 and939, and 3 amino acids were in variations, compared with the PK-01 strain at the sites of 191, 371 and 376;and there were 25 amino acids in variation, compared with the ON1 strain. 10-day-old specific pathogen-free embryonated chicken eggs were inoculated via the Chorioallantoic membrane(CAM) with the W strain. Within 6 days post infection, the EE ratios and the EE indices were calculated. The results showed that, compared with control group, the W strain vaccinated group had the EE ratios in significant difference on the 3 rd day post infection(P<0.05), and in very significant difference on the 6 th day post infection(P<0.01). Therefore, the ratios and indices of EE might be used as important indicators of FAdV-4 causing dwarfism in chicken embryo.

关 键 词：血清4型禽腺病毒 Hexon基因 EE值 EE指数 

分 类 号：S852.65[农业科学—基础兽医学]