畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (3): 451-457.doi: 10.11843/j.issn.0366-6964.2014.03.015

• 遗传繁育 • 上一篇    下一篇

山东五大地方鸡禽白血病病毒主要流行亚型鉴定及其分离株gp85基因的分子特征分析

陈静1,2,王波1,王海明1,程合刚1,武星辰1,孙淑红1*   

  1. (1.山东农业大学动物医学院,泰安 271018;2.江苏省动物预防医学重点实验室,扬州 225009)
  • 收稿日期:2013-09-25 出版日期:2014-03-23 发布日期:2014-03-23
  • 通讯作者: 孙淑红,E-mail: sunshuhong@sdau.edu.cn
  • 作者简介:陈静(1987-),女,河北唐山人,硕士生,主要从事家禽肿瘤病研究,E-mail:jingxiaying2006@163.com
  • 基金资助:

    国家公益性行业(农业)科技专项(201203055);江苏省动物预防医学重点实验室开放课题

Identification of Main Epidemic Subtypies of Avian Leukosis Virus in Five Local Breeds in Shandong Province and Molecular Characteristics of Their gp85 Gene

CHEN Jing1,2,WANG Bo1,WANG Hai-ming1,CHENG He-gang1,WU Xing-chen1,SUN Shu-hong1*   

  1. (1.College of Veterinary Medicine,Shandong Agricultural University,Tai’an 271018, China;2.Key Lab of Preventive Veterinary Medicine of Jiangsu Province, College of Veterinary Medicine,Yangzhou University, Yangzhou 225009, China)
  • Received:2013-09-25 Online:2014-03-23 Published:2014-03-23

摘要:

通过对山东五大地方鸡(寿光鸡、芦花鸡、百日鸡、琅琊鸡、鲁西斗鸡)不同亚型禽白血病病毒(ALV)分离株鉴定及gp85基因序列分析,探究山东五大地方鸡ALV的主要流行亚型及gp85基因分子特征。分别将寿光鸡、百日鸡、芦花鸡的无菌抗凝血和琅琊鸡、鲁西斗鸡的卵白接种CEF培养9~10 d,常规方法提取感染细胞的cDNA,用针对J亚型ALV(ALV-J)gp85基因(924 bp)的引物和A~J(含囊膜蛋白env基因,2 400 bp)的群引物进行PCR扩增、克隆、测序并分析其特点。本研究发现,2009—2011年间,山东五大地方鸡均存在ALV-J感染,5个ALV-J分离株gp85基因之间的相似性为98.4%~99.9%,与原型毒株HPRS-103相似性为97.8%~98.3%;均扩出内源性ALV(ALV-E)片段,与不能产生传染性病毒粒子的ev-1、ev-3、ev-6毒株相似性最高,推测为鸡基因组所携带;同时还发现,琅琊鸡、芦花鸡存在A亚型ALV(ALV-A)的感染。研究结果表明,2009—2011年间的山东五大地方鸡已经普遍存在ALV-J,两大品系中存在ALV-A和ALV-J共感染,同时ALV-E的片段广泛存在,这一结论揭示出山东地方鸡的禽白血病净化工作更加具有挑战性。

Abstract:

In order to identify different subtypies of avian leukosis virus (ALV) and analyze the sequence of gp85 gene,five ALV strains isolated from local chicken breeds in Shandong Province (Shouguang Chicken,Luhua Chicken,Bairi Chicken,Langya chicken,Luxi cock fighting) were amplified and sequenced.The prevailing subgroup of ALV and molecular characteristic of gp85 gene were identified.The sterile anticoagulant blood of Shouguang Chicken,Luhua Chicken,Bairi Chicken and the egg albumen from Langya chicken,Luxi cock fighting were inoculated onto CEF to culture nine to ten days.Proviral DNA was processed routinely.The gp85 and env genes were amplified by using primers subgroup J of ALV (ALV-J) gp85 (924 bp) and A-J group primers (encoding envelope protein containing env gene,2 400 bp).The specific products of PCR amplification were cloned,sequenced and analyzed.The study found that the five groups of local chickens from Shandong Province were infected with ALV-J during 2009 and 2011.The homology of the five ALV-J isolates was between 98.4% and 99.9%.Comparing with prototype strains HPRS-103,the homology was ranged from 97.8% to 98.3%.All of the five endogenous ALV (ALV-E) fragments were amplified,which has a great agreement with ev-1,ev-3,and ev-6 strains that cannot produce infectious virion.It is presumed that these fragments come from the chicken genome.Meanwhile,we also found that Langya and Luhua chicken were infected with subgroup A of ALV (ALV-A).This study showed that ALV-J existed in five groups of local chicken breeds in Shandong Province from 2009 to 2011.The co-infection of ALV-A and ALV-J was discovered in Langya chicken and Luhua chicken,while ALV-E fragments existed in all lines of five local chickens,which reveals that the purification work of ALV for local chicken breeds in Shandong Province will face more challenge.

中图分类号: