传染性喉气管炎病毒gD基因重组禽痘病毒的制备、纯化及初步鉴定  

Preparation,Purification and Preliminary Identification of Recombinant Fowl Pox Virus Expressing Infectious Laryngotracheitis Virus Glycoprotein D Gene

作　　者：杨明凡[1] 崔保安[1] 陈红英[1] 王学斌[1] 魏战勇[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002

出　　处：《中国生物制品学杂志》2009年第4期351-353,共3页Chinese Journal of Biologicals

基　　金：河南省教育厅自然科学研究计划项目(2008A230008).

摘　　要：目的制备传染性喉气管炎病毒(ILTV)gD基因重组禽痘病毒,并进行纯化及初步鉴定。方法将含有ILTV河南长葛株gD基因的禽痘病毒转移载体转染禽痘病毒感染的鸡胚成纤维细胞,采用蓝色蚀斑筛选纯化重组禽痘病毒,并经PCR及间接免疫荧光试验(IFA)进行鉴定。结果经过5轮蚀斑筛选纯化,获得1株重组禽痘病毒rFPV-ILTV-gD,其PCR产物经琼脂糖凝胶电泳,可见约1400bp的ILTVgD特异性条带。IFA检测显示重组禽痘病毒中的ILTVgD基因得到了表达。结论已成功制备并纯化了ILTVgD基因重组禽痘病毒,为预防ILT提供了新的途径。Objective To prepare,purify and preliminarily identify the recombinant fowl pox virus（FPV）expressing infectious laryngotracheitis virus（ILTV）glycoprotein D（gD）gene. Methods The recombinant fowl pox virus transfer vector pSY681-gD-LacZ carrying gD gene of ILTV HNCG strain was transfected to chick embryo fibroblast（CEF）infected with FPV-017 strain,then recombinant FPV was purified by blue plaque screening and identified by PCR and indirect IFA. Results After purification by 5 cycles of blue plaque screening,a recombinant FPV strain rFPV-ILTV-gD was obtained. The specific ILTV gD band at a length of about 1 400 bp was observed on the agarose gel electrophoretic profile of PCR product of rFPV-ILTV-gD,and IFA showed the expression of ILTV gD. Conclusion The recombinant FPV expressing ILTV gD gene was successfully prepared and purified,which provided a novel route for the prevention of ILT.

关 键 词：传染性喉气管炎病毒 GD基因 重组禽痘病毒 

分 类 号：R562.11[医药卫生—呼吸系统] R392-33[医药卫生—内科学]