摘要
以杆状病毒为载体，在昆虫Ｓｆ９细胞中表达了马立克病病毒的与病毒中和反应密切相关的囊膜糖蛋白Ｂ抗原基因（ｇＢ）。用３５Ｓ－蛋氨酸标记重组杆状病ｇＢ－ＡｃＮＰＶ感染细胞溶解物和抗ＭＤＶｇＢ单克隆抗体１ＡＮ８６作免疫沉锭反应，显示出分子量为９８ｋＤ、５５ｋＤ和４９ｋＤ的３条ＭＤＶｇＢ特异性蛋白质带，与天然ＭＤＶｇＢ的１００ｋＤ、６０ｋＤ和４９ｋＤ糖蛋白在大小和带型上类似。在标记过程中加入能抑制蛋白质Ｎ－糖基化的抗菌素ｔｕｎｉｃａｍｙｃｉｎ，或在标记后用能水解糖苷键的Ｎ－内切糖苷酶Ｅｎｄｏ－Ｈ及ｇｌｙｃａｎａｓｅ酶处理标记细胞溶解物，然后再作免疫沉淀反应。结果表明，在昆虫细胞中表达的重组ＭＤＶｇＢ确能象ＭＤＶ感染的鸡成纤维细胞中的天然ｇＢ那样糖基化。二者糖基化的程度及方式略有差别，但亦非常相似。抗重组ＭＤＶｇＢ鸡血清能象抗ＭＤＶ血清那样与ＭＤＶ感染细胞在荧光抗体试验中呈阳性反应，但未能表现病毒中和作用，其原因有待进一步研究。
y using baculovirus AcNPV as a vector,the Marek's disease virus(MDV)gB gene,which was closely related to virus-neutralization, was expressed in the insect cell line Sf9 cells.In the immunoprecipitation of 35S-methionine labeled recombinnt gB-AcNPV infected Sf9cell lysates,both anti-MDV gB monoclonal antibody and anti-MDV chicken serarecognized 3 MDV-specific protein bands of 98kD, 55kD ,and 49kD. which were very similarto the native MDV gB of 100kD, 60kD and 49kD in the sizes and patterns. Treatment ofgB-AcNPV infected Sf9 cells with tunicamycin during the labeling or with N-glycanase andEndo-H after labeling for immunoprecipitation indicated that the recombinant gB(r-gB)wasglycosylated in insect cells as the native gB in the MDV-infected chink embryo fibroblast(CEF)ell system. The r-gB and native gB were very similar although there was some differences between them in the degree and sites for glycosylation. The chicken sera against the r-gB gave positive reaction to MDV-CEF in fluorescence antibody test but failed to show virus-neutralizing activity as anti-MDV chicken sera did, the possible reasons were discussed.
出处
《病毒学报》
CAS
CSCD
北大核心
1994年第1期24-32,共9页
Chinese Journal of Virology
基金
国家自然科学基金
关键词
马立克病病毒
糖蛋白B抗原
载体
Marek's disease virus,Glycoprotein B gene, Baculovirus vector,ImmunoprecipitationAvian Disease and Oncology Laboratory,USDA
