【作者】 成子强； 哈斯阿古拉； 张利； 赵振华；

【Author】 CHENG Zi - qiang~1,Hasiagula~2,ZHANG Li~1,ZHAO Zhen - hua~3 (1.Animal Science and Technology College of Shandong Agricultural University,Taian271018,China;2.The Center of Biology Engineering of Inner Mongolia University,Huhhot010021,China;3.Animal Science and Medicine College of Inner Mongolia Agricultural University, Huhhot010018,China)

【机构】 山东农业大学动物科技学院； 内蒙古大学 生命科学学院生物工程中心； 内蒙古农业大学 动物科学与医学学院；

【摘要】 内蒙古某肉种鸡场40周龄种鸡发生肿瘤性疾病,死淘率15%;经病理组织学检查,增生的肿瘤为典型的髓细胞瘤,因此怀疑此病是J亚群白血病。取病鸡肝电镜观察,在肝细胞胞浆膜下见有圆形的类病毒粒子存在;将肝病料处理后接种于鸡胚成纤维细胞(CEF),经电镜观察,可见正出芽的病毒粒子。为进一步确诊,以ALV-J原型株HPRS-103囊膜gp85基因为基础设计特异性引物,以感染的CEF基因组DNA为模板,体外扩增(PCR)gp85基因,结果获得了924bp的相应片段。将PCR产物进行分子克隆和序列测定,结果表明,内蒙古地区ALV-J gp85蛋白的推测氨基酸与ALV-J原型株HPRS-103、山东SD9901株、美国ADOL-R5-4株及内源性病毒EAV-HP的gp85蛋白的氨基酸同源性分别为97.40%、95.13%、86.69%和85.06%,其中与HPRS-103的同源性最高,与EAV-HP同源性最低。因此可以确定本研究所克隆的基因为ALV-J gp85基因,暂命名为ALV-J-NM8761。更多还原

【Abstract】 Tumor disease was discovered from broiler breeder of 40 weeks - old in a farm of Inner Mongolia with the culled rate of 15%.Histologic examination of tumor - containing tissues showed lesions typical of myelocytomatosis.So this tumor disease was suspected of avian leucosis virus subgroup J(ALV - J).Using tissue homogenate of liver of chickens having marked lesion of myelocytomatosis, inoculated SPF CEF,culturing 7 days.Some CEFs were observed under electronic microscope.The result was that the form of buding virus were seen in cultured CEFs.In order to attest this virus was ALV -J,a pair of primer was designed on the base of HPRS - 103’ gp85 gene and succeed amplified this genein vitro.This four strains viruses were named NM8761,NM8762,NM9875 and NM9876 respectively. Cloned and sequenced NM8761 strain gp85 gene’PCR product,and compared the predicted amino acid with those of HPRS - 103,SD9901,ADOL -R5 -4 and EAV - HP.The results indicated that the pre - dicted amino acid sequences of NM8761,SD9901, ADOL - R5 -4 and EAV - HP gp85 protein had 97.40%,95.13%,86.69%and 85.06%homology,respectively.Among them,the highest homology is HPRS - 103,and the lowest homology is EAV - HP.It is the reason that the gene that amplifiedin vitrois ALV - J virus gene,and it is not avian endogenous retrovirus.The virus of NM8761 strain would be the base for the further research.This paper also confirm that the PCR is a specific and sensitive method for test ALV - J.更多还原