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  

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

作　　者：陈静[1,2] 王波[1] 王海明[1] 程合刚[1] 武星辰 孙淑红[1] 

机构地区：[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

出　　处：《畜牧兽医学报》2014年第3期451-457,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：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

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9~10d,常规方法提取感染细胞的cDNA,用针对J亚型ALV(ALV-J)gp85基因(924bp)的引物和A^J(含囊膜蛋白env基因,2 400bp)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。In order to identify different subtypies of avian leukosis virus(ALV)and analyze the sequence of gp85gene,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 gp85and env genes were amplified by using primers subgroup J of ALV(ALV-J)gp85(924bp)and A-J group primers(encoding envelope protein containing env gene,2 400bp).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 2009and 2011.The homology of thefive 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-6strains 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 2009to 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.

关 键 词：地方鸡 禽白血病病毒(ALV) GP85基因 分子特征 

分 类 号：S852.659.3[农业科学—基础兽医学]